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.     

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.     

Effects of early weaning strategies on growth,survival and digestive enzyme activities in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis> L.) larvae

The effects of weaning strategies of cobia (Rachycentron canadum L.) larvae to commercial microdiets, either from rotifers or from Artemia, on growth, survival and enzymatic digestive capacity, were investigated. In the first experiment, cobia larvae were weaned from rotifers by co-feeding with a microdiet (Otohime) from 8, 13 or 20 days post-hatching (dph). The larvae in the control treatment were fed rotifers (2–12 dph), Artemia nauplii from 7 dph, and co-fed with the microdiet from 20 dph. In the second experiment, the larvae were weaned from Artemia, which was fed to the larvae from 7 dph, by co-feeding with a microdiet (NRD) from 8, 13 or 18 dph. The larvae in control treatment were fed rotifers, then Artemia to the end of the experiment (28 dph). Weaning of cobia larvae onto a microdiet directly from rotifers significantly reduced growth, survival and digestive capacity of the larvae and did not lead to larval acceptance of the microdiet, compared to those weaned from Artemia in the first experiment. Early weaning of cobia larvae onto NRD microdiet (on 8 or 13 dph) from Artemia in the second experiment also reduced growth, survival rate and gut maturation index, compared to those fed live feed. With available microdiets, weaning of cobia larvae could start from Artemia at around 18 dph in order to obtain comparable growth, survival and gut maturation to larvae fed live feed.     

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.     

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.     

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.     

Development of digestive enzymes in larvae of Mayan cichlid Cichlasoma urophthalmus

The development of digestive enzymes during the early ontogeny of the Mayan cichlid (Cichlasoma urophthalmus) was studied using biochemical and electrophoretic techniques. From yolk absorption (6?days after hatching: dah), larvae were fed Artemia nauplii until 15?dah, afterward they were fed with commercial microparticulated trout food (45% protein and 16% lipids) from 16 to 60?dah. Several samples were collected including yolk-sac larvae (considered as day 1 after hatching) and specimens up to 60?dah. Most digestive enzymes were present from yolk absorption (5?C6?dah), except for the specific acid proteases activity (pepsin-like), which increase rapidly from 8?dah up to 20?dah. Three alkaline proteases isoforms (24.0, 24.8, 84.5?kDa) were detected at 8?dah using SDS?CPAGE zymogram, corresponding to trypsin, chymotrypsin and probably leucine aminopeptidase enzymes, and only one isoform was detected (relative electromobility, Rf?=?0.54) for acid proteases (pepsin-like) from 3?dah onwards using PAGE zymogram. We concluded that C. urophthamus is a precocious fish with a great capacity to digest all kinds of food items, including artificial diets provided from 13?dah.     

Weaning requirements of larval mulloway,Argyrosomus japonicus

Mulloway (Argyrosomus japonicus) is an emerging aquaculture species in Australia, but there is a need to improve the production technology and lower costs, including those associated with larval rearing and live feeds. Three experiments were conducted to determine appropriate weaning strategies from live feeds, rotifers (Brachionus plicatilis) and Artemia, to cheaper formulated pellet diets. Experiment 1 examined the effects of feeding Artemia at different levels [0%, 50% or 100% ration of Artemia fed from 18 days after hatching (dah); based on current hatchery protocols] and a pellet diet from two larval ages (14 or 23 dah). In addition, rotifers were supplied to larvae in all treatments for the duration of the experiment (14–29 dah), at which time all larvae were successfully weaned onto the pellet diet. No significant (P>0.05) differences existed between the growth of fish fed a 50% and 100% ration of Artemia; however, fish fed a 0% ration of Artemia had significantly (P<0.05) reduced growth. The time of pellet introduction had no significant (P>0.05) effects on the growth of larvae. Experiments 2 and 3 were designed to determine the size [total length (TL), mm] at which mulloway larvae selected Artemia equally or in preference to rotifers, and pellet (400 μm) equally or in preference to Artemia respectively. Each day, larvae were transferred from a holding tank to experimental vessels and provided with rotifers (2 mL^?1), Artemia (2 mL^?1) or a combination of rotifers (1 mL^?1) and Artemia (1 mL^?1) (Experiment 2), and Artemia (2 mL^?1), a pellet diet or a combination of Artemia (1 mL^?1) and a pellet diet that was broadcast every 15 min (Experiment 3). After 1 h, a sub‐sample of larvae was randomly selected from each replicate vessel (n=5) and the gut contents were examined under a light microscope. Mulloway larvae began selecting Artemia equally to rotifers at 5.2 ± 0.5 mm TL and selected pellets equally to Artemia at 10.6 ± 1.8 mm TL. Our results have led to the establishment of weaning protocols for larval mulloway, which optimize larval growth while reducing feed cost by minimizing the amount of Artemia used during production.     

Optimum time for weaning South African <Emphasis Type="Italic">Scylla serrata</Emphasis> (Forskål) larvae from rotifers to <Emphasis Type="Italic">Artemia</Emphasis>

To determine the optimum time at which to wean Scylla serrata larvae from rotifers onto Artemia two experiments were conducted, approximately 1 month apart, using larvae from two different female crabs. In the first experiment, the larvae in three treatment groups, with nine replicates each, were fed rotifers for the first 8 days after hatching. Artemia were introduced on days after hatch (DAH) 0 – during the first zoeal instar (treatment R + A); on DAH 4 – during the second zoeal instar (treatment R4A); on DAH 8 – during the third zoeal instar (treatment R8A). In a control (ROT) larvae were fed with rotifers exclusively for 18 days until the completion of metamorphosis to megalopa. In the second experiment, the same four feeding schedules as in experiment 1 were used with an additional group of larvae (treatment AC) that were fed only on Artemia throughout the rearing period. Similar results were recorded in the two experiments. Larvae in treatments R + A and R4A performed significantly better than those in treatments R8A, ROT and AC. This was particularly evident when examining the proportion of zoeae which successfully completed metamorphosis to megalopa. Poor performance of larvae in treatments AC and ROT implied that rotifers are needed as a first food, but that rotifers alone do not fill the nutritional requirements of S. serrata larvae. Poor performance of larvae in treatment R8A suggested that the diet should be supplemented with Artemia before the end of the zoea 3 stage.     

Proteolytic Activity in California Halibut Larvae (Paralichthys californicus)

The digestive tract of many marine fish larvae undergoes numerous morphological and functional changes during ontogeny that can substantially influence larval survival under culture conditions. Increasing our knowledge of the digestive capacity and nutritional requirements of the larvae of new candidate species for aquaculture will aid in the development of optimal feeding protocols and greatly improve production under hatchery conditions. In this study, we assess the proteolytic capacity of California halibut (Paralichthys californicus) larvae using biochemical and histological analyses. Newly hatched larvae were reared in a semiclosed recirculating system and fed with highly unsaturated fatty acid (HUFA)–enriched rotifers from hatching until 19 d posthatch (dph) and HUFA‐enriched Artemia nauplii thereafter. Total and specific activity of trypsin and leucine‐aminopeptidase (LAP) and acid and alkaline protease activities were assessed throughout development using spectrophotometric techniques. Trypsin‐like activity and LAP and alkaline protease activities were detected shortly after hatching and before the opening of the mouth. Acid protease activity was not detected until 36–40 dph, concomitant with the development of the gastric glands. The specific activity of trypsin and LAP showed two distinct peaks at 8 and 20 dph. The second peak coincided with the shift from rotifers to Artemia. Hence, newly hatched California halibut larvae possess alkaline proteolytic activity before first feeding. Based on the digestive capacity evaluated in this study and the timing of the development of the functional stomach, we propose that California halibut can be adequately weaned to formulated microdiets around 36 dph.     

Arachidonic acid requirements in larval summer flounder, Paralichthys dentatus

The arachidonic acid (20:4n-6,AA) requirements of larval summer flounder weredetermined for the rotifer- and Artemia-feeding stages. Experimental emulsionscontained adequate n-3 highly unsaturated fattyacid (HUFA) ratios and emulsion levels of AAwere set at 0, 3, 6, 9, and 12% (AA0, AA3,AA6, AA9, and AA12). Examination of fatty acidlevels in live feeds and larval tissuesconfirmed the physiological incorporation offatty acids relative to dietary levels. In thefirst experiment, survival, growth, andsalinity tolerance (2-h in 70) were measuredat 18 days after hatch (dah) after feeding thelarvae the various levels of AA. Larvae fedAA6-enriched rotifers were better able tosurvive the salinity tolerance test. AAenrichment up to 12% had no effect on growthand survival. In the second experiment, larvaewere fed AA0- or AA6-enriched rotifers until 23dah, followed by unenriched 24- and 48-h Artemia nauplii until 32 dah. These larvaethen were subdivided and fed AA-enriched Artemia from 33-45 dah. At the end of thisexperiment, larvae fed AA6-enriched rotifershad higher survival, increased growth, andsurvived better in the salinity tolerance test(2-h in 80). The enrichment of Artemiadid not have any effect on these variables.Thus, the provision of AA6-enriched rotifers(10 mg AA g^–1 DW) early in larvaldevelopment may serve to enhance larval stresstolerance at the rotifer stage while alsoincreasing larval survival, growth, and stresstolerance later in the Artemia stage.     

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.     

The changes in the biochemical compositions and enzymatic activities of rotifer (<Emphasis Type="Italic">Brachionus plicatilis</Emphasis>, Müller) and <Emphasis Type="Italic">Artemia</Emphasis> during the enrichment and starvation periods

The changes in the biochemical compositions and enzymatic activities of rotifer (Brachionus plicatilis) and Artemia, enriched and stored at 4°C temperature, were determined. The total starvation period was 16 h and samples were taken at the end of the 8th and 16th hours. In present study, the rotifer and nauplii catabolized a large proportion of the protein during the enrichment period. Lipid contents of both live preys increased during the enrichment period and decreased in nauplii and metanauplii throughout the starvation period but lipid content of the rotifer remained relatively constant during the starvation period. The changes observed in the amino acid compositions of Artemia and the rotifer were statistically significant (P < 0.05). The conspicuous decline the essential amino acid (EAA) and nonessential amino acid (NEAA) content of the rotifer was observed during the enrichment period. However, the essential amino acid (EAA) and nonessential amino acid (NEAA) contents of Artemia nauplii increased during the enrichment period. The unenriched and enriched rotifers contained more monounsaturated fatty acid (MUFAs) than polyunsaturated fatty acid (PUFAs) and saturated fatty acids (SFA). However, Artemia contained more PUFAs than MUFAs and SFA during the experimental period. A sharp increase in the amounts of docosahexaenoic acid (DHA) during the enrichment of the rotifer and Artemia nauplii was observed. However, the amount of DHA throughout the starvation period decreased in Artemia metanauplii but not in Artemia nauplii. Significant differences in tryptic, leucine aminopeptidase N (LAP), and alkaline phosphatase (AP) enzyme activities of Artemia and rotifer were observed during the enrichment and starvation period (P < 0.05). The digestive enzymes derived from live food to fish larvae provided the highest contribution at the end of the enrichment period. In conclusion, the results of the study provide important contributions to determine the most suitable live food offering time for marine fish larvae. Rotifer should be offered to fish larvae at the end of the enrichment period, Artemia nauplii just after hatching and before being stored at 4°C, and Artemia metanauplii at the end of the enrichment and throughout the starvation period.     

Histological development of digestive tract in discus, <Emphasis Type="Italic">Symphysodon</Emphasis> spp. larvae

The present study provides information on the histomorphological development of digestive system of discus, Symphyosodon spp., larvae during the first month of life. Discus larvae are altricial at hatching, with an undifferentiated digestive tract and a large yolksac, which is completely consumed within 7 days. The mouth opens 3 days after hatching (DAH) and the larvae starts feeding on AF Artemia at 4 DAH when offered. At 3 DAH the digestive tract is differentiated with distinct esophagus, stomach anlage, and mid- and hindguts. At 5 DAH, discus larvae is an active feeder, equipped with partly developed jaws and ossified gill arches and an inflated swim bladder. The liver and pancreas are present and supranuclear inclusion vacuoles (SIV) appear in the hindgut for the first time. Gastric glands in stomach were first observed 7 DAH and proliferated by 11–13 DAH. SIV were a common feature in the midgut and hindgut epithelium until 15–23 DAH. Therefore, exclusive use of artificial diets should be postponed until 2–3 weeks after hatching.     

Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: Effects on growth performance and digestive enzyme activities

In this study, the influence of commercial probiotic, Lactobacillus spp., supplementation was investigated on growth parameters and digestive enzyme activities in gilthead sea bream, Sparus aurata, during larval development. All experiments were triplicated and designed in three different administrations of probiotic from 3 days after hatching (DAH) concurrently with starting of exogenous feeding. In the first group, probiotic was added to live food (rotifer and Artemia). In the second group, probiotic was supplemented directly to both live food and water. In the third group, probiotic was added directly to water. Also, no probiotic treatment was maintained in control group. Total bacterial counts among probiotic probiotic-supplemented groups were significantly different from total bacterial counts in controls in water and digestive tract of larvae (p < 0.05). The mean of total bacterial counts in control was approximately 4 × 10⁴-fold increased from the experimental groups in the sea water (p < 0.05). Besides, mean digestive enzyme activities of all probiotics treatment groups were significantly different (p < 0.05) with that of the control. Except probiotic water supplementation group, in all treatments, the specific activities of pancreatic and intestinal enzymes were significantly higher (p < 0.05) in larvae to which probiotic had been supplemented by live food and live food with water. Also, S. aurata larvae that had probiotic administered by live food with water demonstrated significant (p < 0.05) increases in both survival (13–105% higher) and specific growth rate (2–9% higher) as compared to controls. As a result, supplementation of probiotic to directly tank water could not significantly increase growth parameters and digestive enzyme activities and therefore, administration of probiotics by this method would not be effective in terms of husbandry parameters and nutritional condition.     

Establishing larval feeding regimens for the Forktail Blenny Meiacanthus atrodorsalis (Günther, 1877): effects of Artemia strain,time of prey switch and co‐feeding period

This study aimed to establish feeding strategies covering the whole larval period of the forktail blenny, Meiacanthus atrodorsalis, based on the standard hatchery feeds of rotifers and Artemia. Three purposely designed experiments were conducted to determine the appropriate times and techniques to transition larvae from rotifers onto Artemia nauplii of a Great Salt Lake (GSL) strain, and a specialty AF strain, as well as subsequent transition onto enriched metanauplii of GSL Artemia. With a 3‐day co‐feeding period, larvae adapted well to a transition from rotifers to newly hatched GSL Artemia nauplii as early as 5 days posthatching (DPH), and as early as 3 DPH when fed the smaller AF Artemia nauplii. However, prolonging the rotifer‐feeding period up to 11 DPH did not negatively affect survival. Larvae fed Artemia nauplii of the AF strain showed 17–21% higher survival, 24–33% greater standard length and body depth, and 91–200% greater dry weight, after 20 days relative to those fed nauplii of the GSL strain. Meanwhile, enriched Artemia metanauplii of the GSL strain were shown to be an acceptable alternative to AF Artemia nauplii for later larvae, producing similar survival and growth when introduced from 8 DPH. Based on our findings, we recommend feeding M. atrodorsalis larvae rotifers as a first food between 0 and 2 DPH, introducing AF Artemia nauplii from 3 DPH, followed by enriched GSL Artemia metanauplii from 8 DPH onward, with a 3‐day co‐feeding period between each prey change.     

Natural carbon stable isotope ratios as indicators of the relative contribution of live and inert diets to growth in larval Senegalese sole (Solea senegalensis)

The relative contributions of live Artemia metanauplii and an inert diet for growth of Senegalese sole larvae and postlarvae were assessed through the analysis of carbon stable isotopes ratios (δ¹³C) in both diets and whole larval tissue. Larvae were reared on four dietary regimes: 100% live prey (rotifers and Artemia), 100% inert formulated diet and two co-feeding regimes of 70:30 and 30:70 ratios of Artemia and inert diet, respectively. Larvae from the live food regime and both co-feeding regimes showed a steep increase in δ¹³C from 10 days after hatching (DAH) as a result of the onset and continuation of Artemia consumption. From 12 DAH fish larvae from all the regimes showed significant isotopic differences as their δ¹³C increased to final asymptotic values of − 15.1, − 15.6 and − 16.3‰ in the live food, 70:30 and 30:70 regimes, respectively. Carbon turnover rates in larvae from both live food and co-feeding regimes were relatively high (0.071 to 0.116 d^− 1) but more than 90% of the observed change in fish tissue isotopic values was accounted for by the retention of carbon in new tissue growth. A two-source, one-isotope mixing model was applied to estimate the nutritional contribution of Artemia and inert diet to postlarvae growth in the co-feeding regimes. At 23 DAH, the relative contribution of live and inert diets to tissue growth in larvae was respectively, 88 and 12% for the 70:30 co-feeding regime and 73 and 27% for the 30:70 co-feeding regime. At 17 DAH, the estimated proportion of tissue carbon derived from the inert diet was higher at 23 and 38% for the 70:30 and 30:70 regimes, respectively. The results suggest that co-feeding regimes in Solea senegalensis larvae may be adjusted to meet ontogenetic changes in the capacity for larvae to utilise inert diets. The contrasting levels of carbon isotope discrimination between diet and tissue in larvae reared on either 100% live feed or 100% inert diet indicate relatively poor utilization of nutrients from the inert diet. The use of isotopic discrimination factors as potential indicators of the digestive physiological performance of a consuming organism in regards to its diet is discussed.     

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.     

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.     

Optimal first feed organism for South African mud crab <Emphasis Type="Italic">Scylla serrata</Emphasis> (Forskål) larvae

It is not known whether rotifers or Artemia nauplii are the best first food for South African mud crab Scylla serrata larvae. In order to test this, larvae were fed with five different test diets. These were rotifers for the first 8 days and newly hatched EG^® type Artemia nauplii (San Francisco Bay) from day 6 onwards (treatment R6A); newly hatched EG^® type Artemia nauplii throughout the rearing period (treatment EG); newly hatched Vinh-Chau strain (Vietnam) Artemia nauplii throughout the rearing period (treatment VC); decapsulated cysts of EG^® type Artemia throughout the rearing period (treatment DECAP); or decapsulated cysts supplemented with low densities of Artemia EG type Artemia nauplii (treatment MIX). Two experiments were conducted approximately 1 month apart using larvae from two different female crabs. Although results showed it is possible to rear S. serrata larvae through metamorphosis on Artemia nauplii exclusively, larval performance (development, survival and successful metamorphosis) was enhanced by the inclusion of rotifers as a first feed.No significant difference in performance was recorded between larvae fed on the two strains of Artemia nauplii. Larvae fed on decapsulated cysts in treatments DECAP and MIX performed poorly, but there were indications that decapsulated cysts and other inert diets may have potential as supplements to live food in the rearing of S. serrata larvae.