Larval rearing and ontogeny of digestive enzyme activities in yellowfin seabream (Acanthopagrus latus, Houttuyn 1782).

The present research was conducted to provide insight into digestive larval capacity in Acanthopagrus latus larvae from hatching up to 30 days after hatching (DAH). Newly hatched larvae were stocked into six 300-L cylindrical polyethylene tanks at a density of larvae 50 larvae/L and reared by means of the green water system using Nannochloropsis oculata (0.5 × 106/mL). After mouth opening, larvae were fed with rotifers (5-16 individual/mL) from 2 to 20 DAH; then, Artemia nauplii (0.5-3.0 individuals/mL) were offered to larvae from 18 to 30 DAH, meanwhile a commercial microdiet was offered to larvae from 25 to 30 DAH. Larval performance in terms of growth and survival, and the assessment of the activity of selected digestive enzymes ontogeny of digestive enzymes activities was evaluated in larvae sampled at 0 (hatching), 7, 15, 22 and 30 DAH. Larvae showed an exponential growth characterized by two different growth stanzas, a first one characterized by slow growth rates comprised between hatching to 15 DAH (4.7 ± 0.2 mm), followed by a period of faster growth rates between 16 and 30 DAH (7.5 ± 0.6 mm). The activities of the brush border (alkaline phosphatase, ALP) and cytosolic (leucine-alanine peptidase, LAP) enzymes, as well as those of the pancreatic ones like total alkaline proteases, bile salt-activated lipase and α-amylase were detected from the mouth opening stage. Total activities of pancreatic and gastric enzymes increased with larval growth showing an enhancement of digestive capacities with larval age and size. The intestinal maturation in A. latus as assessed by the ratio of AP to LAP did not occur as expected by end of the first month of life suggesting the complete establishment of digestive luminal processes may take place at older ages. This study related to the growth patterns and ontogenic changes in activity of pancreatic, gastric and intestinal enzymes in A. latus and their nutritional regulation may be considered as the first step for improving the larviculture, as well as assessing and refining the nutritional requirements during the larval and early juvenile stages of this sparid species.

Suction feeding across fish life stages: flow dynamics from larvae to adults and implications for prey capture.

Suction feeding is thought to be the primary mode of prey capture in most larval fishes. Similar to adult suction feeders, larvae swim towards their prey while rapidly expanding their mouth cavity to generate an inward flow of water that draws the prey into the mouth. Although larvae are known to experience flows with lower Reynolds numbers than adults, it is unclear how the suction-induced flow field changes throughout ontogeny, and how such changes relate to prey capture performance. To address these questions, we determined mouth dimensions and opening speeds in Sparus aurata from first-feeding larvae to adults. We proceeded to develop a computational model of mouth expansion in order to analyze the scaling of suction flows under the observed parameters. Larval fish produced suction flows that were around two orders of magnitude slower than those of adults. Compared with adult fish, in which flow speed decays steeply with distance in front of the mouth, flow speed decayed more gradually in larval fish. This difference indicates that viscous forces in low Reynolds number flows modify the spatial distribution flow speed in front of the mouth. Consequently, simulated predator-prey encounters showed that larval fish could capture inert prey from a greater distance compared with adults. However, if prey attempted to escape then larval fish performed poorly: simulations inferred capture success in only weakly escaping prey immediately in front of the mouth. These ontogenetic changes in Reynolds number, suction-induced flow field and feeding performance could explain a widespread ontogenetic diet shift from passive prey at early life stages to evasive prey as larvae mature.

Long lasting effects of early temperature exposure on the swimming performance and skeleton development of metamorphosing Gilthead seabream (Sparus aurata L.) larvae.

Temperatures experienced during early ontogeny significantly influence fish phenotypes, with clear consequences for the wild and reared stocks. We examined the effect of temperature (17, 20, or 23 °C) during the short embryonic and yolk-sac larval period, on the swimming performance and skeleton of metamorphosing Gilthead seabream larvae. In the following ontogenetic period, all fish were subjected to common temperature (20 °C). The critical swimming speed of metamorphosing larvae was significantly decreased from 9.7 ± 0.6 TL/s (total length per second) at 17 °C developmental temperature (DT) to 8.7 ± 0.6 and 8.8 ± 0.7 TL/s at 20 and 23 °C DT respectively (p < 0.05). Swimming performance was significantly correlated with fish body shape (p < 0.05). Compared with the rest groups, fish of 17 °C DT presented a slender body shape, longer caudal peduncle, terminal mouth and ventrally transposed pectoral fins. Moreover, DT significantly affected the relative depth of heart ventricle (VD/TL, p < 0.05), which was comparatively increased at 17 °C DT. Finally, the incidence of caudal-fin abnormalities significantly decreased (p < 0.05) with the increase of DT. To our knowledge, this is the first evidence for the significant effect of DT during the short embryonic and yolk-sac larval period on the swimming performance of the later stages.

Cloning and molecular ontogeny of digestive enzymes in fed and food-deprived developing gilthead seabream (Sparus aurata) larvae.

We have determined the expression pattern of key pancreatic enzymes precursors (trypsinogen, try; chymotrypsinogen, ctrb; phospholipase A2, pla2; bile salt-activated lipase, cel; and α-amylase, amy2a) during the larval stage of gilthead seabream (Sparus aurata) up to 60days after hatching (dph). Previously, complete sequences of try, cel, and amy2a were cloned and phylogenetically analyzed. One new isoform was found for cel transcript (cel1b). Expression of all enzyme precursors was detected before the mouth opening. Expression of try and ctrb increased during the first days of development and then maintained high values with some fluctuations during the whole larval stage. The prolipases pla2 and cel1b increased from first-feeding with irregular fluctuation until the end of the experiment. Contrarily, cel1a maintained low expression values during most of the larval stage increasing at the end of the period. Nevertheless, cel1a expression was negligible as compared with cel1b. The expression of amy2a sharply increased during the first week followed by a gradual decrease. In addition, a food-deprivation experiment was performed to find the differences in relation to presence/absence of gut content after the opening of the mouth. The food-deprived larvae died at 10dph. The expression levels of all digestive enzymes increased up to 7dph, declining sharply afterwards. This expression pattern up to 7dph was the same observed in fed larvae, confirming the genetic programming during the early development. Main digestive enzymes in gilthead seabream larvae exhibited the same expression profiles than other marine fish with carnivorous preferences in their juvenile stages.

Ontogeny of the digestive tract in sharpsnout sea bream Diplodus puntazzo (Cetti, 1777).

The ontogeny of the digestive tract was studied histologically and histochemically in sharpsnout sea bream Diplodus puntazzo from hatching (0 DAH, Days After Hatching) until day 57 (57 DAH). At hatching, the digestive tract appeared as a histologically undifferentiated straight tube lying dorsally to the yolk sac. When the mouth opened at 3 DAH, the digestive tract was differentiated into buccopharynx, oesophagus, incipient stomach and intestine. The pancreas, liver and gall bladder were also differentiated at this stage and both the bile and pancreatic duct had opened into the anterior intestine. Active feeding began in 50% of larvae at 4 DAH, although permanence of yolk reserves until 7 DAH suggests a period of both endogenous and exogenous feeding. Nutrient absorption was first visible from 5 DAH, as colourless supra- and infranuclear vacuoles in the anterior intestinal mucosa, suggesting a lipid content, as well as supranuclear, eosinophilic vacuoles, containing protein, in the posterior intestinal mucosa. Early caecal development could be detected from 10 DAH, whereas gastric glands appeared at 30 DAH, indicating the transition from larval to juvenile stage and the acquisition of an adult mode of digestion. Goblet cells appeared in the digestive tract of sharpsnout sea bream larvae shortly after first feeding. The mucus content of goblet cells varied with the digestive region and, in the buccal cavity and oesophagus, also with the developmental phase. This study provides knowledge for better husbandry practices in the aquaculture industry, as well as for the implementation of future nutritional studies.