Ontogenetic development of digestive enzymes in yellowtail kingfish Seriola lalandi larvae

The development of digestive enzymes was examined in laboratory-reared yellowtail kingfish larvae from hatching to 36 days after hatching (DAH). The specific activities of amylase, lipase, and alkaline phosphatase showed three distinct phases: a sharp increase in enzyme activity from hatching to the onset of exogenous feeding on 3 DAH, followed by a fluctuation and a general decline toward 18 DAH, and then a period of low activity from 18 to 36 DAH. The total activities of these three enzymes showed a gradual increase from hatching to 18 DAH, followed by a sharp increase toward 36 DAH. In contrast to other enzymes, the specific and total activities of trypsin reached the maximum on 15 DAH and 24 DAH, respectively, and then both activities declined to low levels toward 36 DAH. The dynamics of digestive enzymes corresponded to the anatomical development of the digestive system. The enzyme activities tend to be stable after the formation of gastric glands in the stomach on 15 DAH. The composition of digestive enzymes indicates that yellowtail kingfish is able to digest protein, lipid and carbohydrates at an early stage. However, due to the low level of amylase specific activity after 18 DAH, the carbohydrate component should remain at a low level in formulated diets for fish larvae.     

泥鳅仔稚鱼发育期间消化酶及碱性磷酸酶比活力的变化

研究了泥鳅(Misgurnus anguillicaudatus)从孵化至30 DAH(日龄,Days after hatching)几种消化酶及碱性磷酸酶比活力的变化情况。胃蛋白酶直至30 DAH仍未检出活性。而胰蛋白酶表现出较高的比活力,其比活力在初次摄食之后显著上升,6 DAH达到最大值之后开始显著降低(P<0.05)。脂肪酶与淀粉酶的变化模式相似,在内源性营养向外源性营养转变及仔鱼向稚鱼转变这两个时间段出现两个高峰值。碱性磷酸酶比活力在2-6 DAH显著上升(P<0.05),之后开始下降并趋于平稳。研究表明,泥鳅在仔稚鱼阶段只具有结构性的胃而缺乏分泌细胞的分化。2-6 DAH是泥鳅仔鱼肠道功能迅速发育的阶段,也是向成鱼消化模式转变的一个重要过程。脂肪酶和淀粉酶比活力的持续性表明了泥鳅仔鱼对糖类和脂肪有较好的利用能力。     

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。     

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。     

Changes in digestive enzyme activities during larval development of leopard grouper (Mycteroperca rosacea)

The leopard grouper is an endemic species of the Mexican Pacific with an important commercial fishery and good aquaculture potential. In order to assess the digestive capacity of this species during the larval period and aid in the formulation of adequate weaning diets, this study aimed to characterize the ontogeny of digestive enzymes during development of the digestive system. Digestive enzymes trypsin, chymotrypsin, acid protease, leucine–alanine peptidase, alkaline phosphatase, aminopeptidase N, lipase, amylase and maltase were quantified in larvae fed live prey and weaned onto a formulated microdiet at 31 days after hatching (DAH) and compared with fasting larvae. Enzyme activity for trypsin, lipase and amylase were detected before the opening of the mouth and the onset of exogenous feeding, indicating a precocious development of the digestive system that has been described in many fish species. The intracellular enzyme activity of leucine–alanine peptidase was high during the first days of development, with a tendency to decrease as larvae developed, reaching undetectable levels at the end of the experimental period. In contrast, activities of enzymes located in the intestinal brush border (i.e., aminopeptidase and alkaline phosphatase) were low at the start of exogenous feeding but progressively increased with larval development, indicating the gradual maturation of the digestive system. Based on our results, we conclude that leopard grouper larvae possess a functional digestive system at hatching and before the onset of exogenous feeding. The significant increase in the activity of trypsin, lipase, amylase and acid protease between 30 and 40 DAH suggests that larvae of this species can be successfully weaned onto microdiets during this period.     

Developmental changes in digestive enzyme activity in American shad, <Emphasis Type="Italic">Alosa sapidissima</Emphasis>, during early ontogeny

In order to assess the digestive physiological capacity of the American shad Alosa sapidissima and to establish feeding protocols that match larval nutritional requirements, we investigated the ontogenesis of digestive enzymes (trypsin, amylase, lipase, pepsin, alkaline phosphatase, and leucine aminopeptidase) in larvae, from hatching to 45 days after hatching (DAH). We found that all of the target enzymes were present at hatching, except pepsin, which indicated an initial ability to digest nutrients and precocious digestive system development. Trypsin rapidly increased to a maximum at 14 DAH. Amylase sharply increased until 10 DAH and exhibited a second increase at 33 DAH, which coincided with the introduction of microdiet at 30 DAH, thereby suggesting that the increase was associated with the microdiet carbohydrate content. Lipase increased until 14 DAH, decreased until 27 DAH, and then increased until 45 DAH. Pepsin was first detected at 27 DAH and then sharply increased until 45 DAH, which suggested the formation of a functional stomach. Both alkaline phosphatase and leucine aminopeptidase markedly increased until 18 DAH, which indicated intestinal maturation. According to our results, we conclude that American shad larvae possess the functional digestive system before mouth opening, and the significant increases in lipase, amylase, pepsin, and intestinal enzyme activities between 27 and 33 DAH suggest that larvae can be successfully weaned onto microdiets around this age.     

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.     

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.     

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.     

Ontogenic changes in the digestive enzyme activities and the effect of different starvation duration on the digestive enzyme activities of larval red swamp crayfish (Procambarus clarkii)

This study was performed to determine the effect of starvation and delayed feeding on activities of digestive enzymes and alkaline phosphatase (ALP) of larval red swamp crayfish (Procambarus clarkii), so as to reveal the tolerance to prolonged starvation and the recovery of digestive enzymes after delayed feeding in larval and juvenile P. clarkii. In the control group, activities of trypsin and ALP increased significantly (p < .05) with day‐age and then kept constant at 24 days after hatching (DAH) and 10 DAH, respectively, whereas the activities of amylase and pepsin increased firstly then decreased with day‐age, and the activity of lipase increased firstly then decreased and then increased again during the development period of juvenile P. clarkii (1–31 DAH). In the group with continuous starvation (CS), activities of pepsin and lipase both decreased (p < .05) after fasting, and the activities of pepsin, lipase and trypsin in the groups with delayed feeding all increased (p < .05) and recover to the levels of the control group after food supply. However, the activity of amylase increased (p < .05) in the CS group, and it decreased to normal level after food supply. The ALP activity did not significantly (p > .05) vary after starvation, whereas it decreased in the groups with delayed feeding after 1 day of food supply, and then increased back to the level similar with the control group. Results from this study could provide information for diet preparation and feeding regime in larval and juvenile red swamp crayfish culture.     

Ontogenic development of digestive enzymes in bigfin reef squid (Sepioteuthis lessoniana)

Bigfin reef squid (Sepioteuthis lessoniana) is an economically important exported aquatic animal in Thailand, but little is known about its husbandry, which needs to be clarified. In this study, the ontogenic development of the main digestive enzymes (pepsin, trypsin, chymotrypsin, lipase and amylase) was investigated within 30 days after hatching (DAH). Fertilized eggs of the wild cuttlefish were hatched and then distributed into four replicate aquaria assigned to varying collection times (0, 5, 10, 15, 20, 25 and 30 DAH). The development of pepsin suggests acidic digestion functionality at 5 DAH, and its specific activity was maintained over the studied period. Specific activities of the serine proteases, trypsin and chymotrypsin, fluctuated significantly with similar pattern, and appear to be the key enzymes contributing ontogenic changes in protein catabolism. The specific activity of lipase was relatively high at 0 DAH, dramatically increased by 5 DAH, and then appeared constant until the end of observations, suggesting important role in utilizing the lipid from yolk at hatching, as well as probably in sparing protein utilization. Dramatic increase in amylase specific activity occurred by 10 DAH after which the activity was constant until 30 DAH, suggesting the utilization of carbohydrate as energy evolves comparatively late. Findings from the current study could be used to manage nutritional protocols according to the digestive enzyme patterns, as well as to develop artificial diets for rearing S. lessoniana.     

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.     

Digestive enzyme activity of the red porgy (Pagrus pagrus, L.) during larval development under culture conditions

The ontogenic development of the main digestive enzymes (proteases, amylase and lipase) in the red porgy, Pagrus pagrus, larvae was assayed during the larval development. The green water technique was carried out for larval rearing and whole‐body homogenates were used for enzymatic assays in triplicate. Significant alterations in specific activities of all digestive enzymes measured during the period of this study were mostly related to metamorphosis and weaning. Trypsin‐ and chymotrypsin‐specific activities were first detected on day 3, together with opening of the mouth, and slightly increased until 25 days after hatching (DAH). After this period, the specific activities of these enzymes slightly decreased. Pepsin was first detected on day 28, concurrent with stomach formation, and a sharp increase was observed until 30 DAH. A slight decrease was measured from this date until the end of the experiment. Both amylase and lipase were measured for the first time on days 2 and 4 respectively, and the specific activities of these enzymes showed similar patterns during the first week of the study. Then, slight variations were observed until 30 DAH and while lipase‐specific activity declined, an increase in the specific activity of amylase was found until the end of the experiment. It is concluded that the variations observed in the specific activity of digestive enzymes were related to either metamorphosis, such as the formation of the stomach (28 DAH), or to changes in food composition. The profile of the developmental pattern of the main digestive enzymes detected in P. pagrus is similar to that described for other Sparid species.     

Ontogenic development of the digestive enzymes in common pandora, Pagellus erythrinus, L. larvae

Ontogenic development of some digestive enzymes (proteases, amylase and lipase) in common pandora Pagellus erythrinus larva was assayed during larval development. The green‐water technique was employed for larval rearing, and whole‐body homogenates were used for enzymatic assays in triplicate. Important alterations in specific activities of all digestive enzymes measured during the period of this study were mostly related to metamorphosis and weaning. Mouth opening was observed on day 3 at 2.23±0.01 mm total length synchronously with the first determination of trypsin and chymotrypsin activities. After this date, the specific activities of these slightly increased until 25 days after hatching (DAH), respectively, and then slightly decreased and changed. The pattern determined for pepsin was strongly related to stomach formation on day 25 at 9.72±2.3 mm total length and a sharp increase was found until 30 DAH and then a slight decrease was measured from this date until the end of the experiment. Both amylase and lipase were measured for the first time on days 2 and 4, respectively, and also the specific activities of these showed similar patterns during the first week of the study. Then, slight variations were observed until 30 DAH and while lipase‐specific activity had declined, an increase in the specific activity of amylase was found until the end of the experiment. Finally, it is thought that the variations observed in the specific activity in the profile of digestive enzymes were related to either metamorphosis such as formation of stomach (25 DAH) or to changes in characteristics of food (30 DAH). The pattern of development of the main digestive enzymes found in P. erythrinus is similar to that described in other Sparid species.     

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 Enzymes in California Halibut <Emphasis Type="Italic">Paralichthys californicus</Emphasis> Larvae

California halibut Paralichthys californicus is an important commercial species with high aquaculture potential in Baja California Sur, México. To optimize the feeding process using live prey and/or inert diets, we evaluated alkaline proteases, pepsin, trypsin, chymotrypsin, leucine aminopeptidase, lipase, α-amylase, and acid and alkaline phosphatase activities on starved larvae and larvae fed live prey. Highest activities were observed for alkaline protease, trypsin, chymotrypsin, leucine aminopeptidase, and alkaline phosphatase in feeding larvae than starved larvae on day 4 after hatching. At day 5, a sizeable increase in all enzymatic activities was detected in feeding larvae. Alkaline protease, trypsin, chymotrypsin, and alkaline phosphatase decreases progressively from day 5 until day 18. At day 18, a slight pepsin activity was observed. This was considered an indicator of the start of digestive system maturation. We concluded that total enzymatic equipment for this species is complete between day 18 and 30 after hatching. Based on this evidence, early weaning from live prey to inert feed would be possible at this time.     

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.     

Screening of enzyme activity for assessing the condition of larvae in the seven-band grouper <Emphasis Type="Italic">Epinephelus septemfasciatus</Emphasis> and devil stinger <Emphasis Type="Italic">Inimicus japonicus</Emphasis>

We conducted screening tests to determine whether enzyme activity is a suitable biomarker for assessing the physiological condition of marine fish larvae. The rearing experiments consisted of three trials, of which two were conducted using the seven-band grouper Epinephelus septemfasciatus for a period of 5 days after hatching (DAH), and one was conducted using the devil stinger Inimicus japonicus for 10 DAH. The trials were conducted under three different rearing-tank environments (shallow tank, intermediate tank, deep tank) in a water volume of 100 l and an aeration rate of 50 ml/min. We determined survival, surface death, growth, and enzyme activities (trypsin, esterase, and alkaline phosphatase). The highest survival rates and lowest surface deaths in both species occurred among the larvae grown in the deep tank. There was a significant and negative correlation between survival at 5 DAH and alkaline phosphatase activity at 0 DAH in the seven-band grouper. The same correlation was found between survival at 10 DAH and trypsin and alkaline phosphatase activity at 1 DAH in the devil stinger. Based on these results, we conclude that the activity of a specific enzyme is a candidate for assessing the physiological condition of marine fish 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.     

Functional changes in digestive enzyme activities of meagre (Argyrosomus regius; Asso, 1801) during early ontogeny

The ontogenesis of main pancreatic and intestinal enzymes was investigated in the recent promising Mediterranean candidate species of meagre, Argyrosomus regius, during larval development until 40 days after hatching (DAH). The green-water technique was carried out for larval rearing. Whole-body homogenates were used for enzymatic analysis in larvae younger than 15 DAH; after this date, older larvae were dissected into two segments as pancreatic and intestinal segment. Trypsin was detected as early as hatching and sharply increased concurrently with age and exogenous feeding 15 DAH, but constant decline was observed until the end of experiment. Amylase was determined at 2 DAH and sharply increased 10 DAH. Then, slight decreases were found between 10 and 15 DAH, and then slow alterations were continued until the end of the experiment. Lipase was firstly measured on day 3; then, sudden decline was observed between 20 and 25 DAH. After this date, slow fluctuations were maintained until the end of the experiment. Pepsin was firstly assayed 15 DAH related to gastric gland secretion and sharply increased 30 DAH. Then, it slowly varied until end of the experiment. Enzymes of brush border membranes, alkaline phosphatase and aminopeptidase N showed similar pattern on specific activities during the first 10 days. Thereafter, while specific activity of alkaline phosphatase slightly decreased 15 and fluctuated until 20 DAH, aminopeptidase N activity slowly increased 20 DAH. Then, activity of alkaline phosphatase and aminopeptidase N constantly increased 30 DAH, indicating maturation of the intestinal digestive process, and also, these activities continued to slowly increase until the end of the experiment. The specific activity of cytosolic peptidase, leucine–alanine peptidase, smoothly increased on day 8, then fluctuated until 15 DAH. After this date, in contrast to enzymes of brush border membranes, it sharply decreased 25 DAH and continued to gradually decline until the end of the experiment. These converse expressions were indicative of a maturation of enterocytes and the transition of an adult mode of digestion.