Effect of increased rearing temperature on digestive function in cobia early juvenile.

The present study is focused to elucidate the main characteristics of the digestive function of this carnivorous fast-growing fish living at high temperatures. With this aim, we have examined the effects of an increased temperature from 30 to 34 °C on the daily pattern of gastrointestinal pH, enzymatic proteolytic digestive activity and the feed transit time in early juveniles of cobia (Rachycentron canadum), a species living in tropical and subtropical waters with an increasing aquaculture production. Fish were fed two meals a day. Gastric luminal pH was permanently acidic (mean pH values: 2.76-4.74) while the intestinal pH increased from neutral/slightly acidic to slightly alkaline when the digesta was present, with an increasing alkalinity from proximal to distal intestine (mean pH values: 6.05 to 7.69). The temperature did not affect the gastric pH but a slightly higher acidity was induced in the intestine at 34 °C. Pepsin activity showed a daily rhythm at 30 °C with maximum in the middle of the light period, while at 34 °C some hourly changes coinciding with feed adding without a clear daily trend during the 24-h period were observed. The trypsin activity exhibited a daily rhythm at both temperatures with an increase after morning feeding to reach a maximum several hours later. Average pepsin activity during the daily cycle was slightly higher at 34 °C (6.1 and 7.3 U mg-1 BW at 30 and 34 °C respectively), but values were significantly different only at 8 and 24 h after the morning meal. Similarly, the trypsin activity was significantly affected by the temperature only at 8 and 16 h after the morning meal, but daily activity averages were similar (1.20 and 1.29 U g-1 BW at 30 and 34 °C respectively). The partial transit rates of the first meal in the stomach for each period inter-samplings were higher during the first 4-h period and decreased progressively along the rest of the 24-h cycle at both temperatures, but no significant differences were detected at 30 °C. In addition, the transit was notably faster at 34 °C particularly during the first 8 h after feeding, with rates between 100 and 65% of total volume displaced (intake or released) during each 4-h period. In the intestine the transit rate was relatively constant and similar at both temperatures during 12 h after feeding. Then the rates remained very low during the following 12 h. Residence time of the first meal was longer at 30 than at 34 °C, particularly in the stomach (12 h:02 min vs 4 h:54 min respectively). In the intestine the difference was not so large (8 h:18 min vs 6 h:24 min respectively). In a parallel study under same conditions, cobia reared at 30 °C grew faster and showed a more favorable feed conversion ratio than those at elevated temperature (34 °C). The present results indicate that at 34 °C, a subtle increase of proteolytic activity cannot compensate for the faster gut transit rate. Therefore, 30 °C is more appropriate temperature for the early on-growing of cobia because at higher temperatures the digestion efficiency decrease being one of the causes for a lower growth.

The circadian transcriptome of marine fish (Sparus aurata) larvae reveals highly synchronized biological processes at the whole organism level.

The regulation of circadian gene expression remains largely unknown in farmed fish larvae. In this study, a high-density oligonucleotide microarray was used to examine the daily expression of 13,939 unique genes in whole gilthead sea bream (Sparus aurata) larvae with fast growth potentiality. Up to 2,229 genes were differentially expressed, and the first two components of Principal Component Analysis explained more than 81% of the total variance. Clustering analysis of differentially expressed genes identified 4 major clusters that were triggered sequentially, with a maximum expression at 0 h, 3 h, 9-15 h and 18-21 h zeitgeber time. Various core clock genes (per1, per2, per3, bmal1, cry1, cry2, clock) were identified in clusters 1-3, and their expression was significantly correlated with several genes in each cluster. Functional analysis revealed a daily consecutive activation of canonical pathways related to phototransduction, intermediary metabolism, development, chromatin remodeling, and cell cycle regulation. This daily transcriptome of whole larvae resembles a cell cycle (G1/S, G2/M, and M/G1 transitions) in synchronization with multicellular processes, such as neuromuscular development. This study supports that the actively feeding fish larval transcriptome is temporally organized in a 24-h cycle, likely for maximizing growth and development.

Effects of green macroalgal blooms on the meiofauna community structure in the Bay of Cádiz.

The effect of macroalgal blooms on the abundance and community structure of intertidal sediment meiofauna was studied using an in situ enclosure experiments (Bay of Cádiz, Spain). Meiofaunal abundance (3500-41,000 ind 10 cm⁻²) was three to sevenfold higher in the presence of macroalgae. Nematoda were the dominant taxon both in Control (52-82%) and Macroalgae plots (92-96%), followed by Harpacticoida Copepoda and Ostracoda. Non-metric Multi-Dimensional Scaling (MDS) analysis clearly separated the meiofaunal community from Control and Macroalgae plots. Organic matter, organic carbon, total nitrogen, chlorophyll a and freeze-lysable inorganic nutrients were higher in Macroalgae plots, and were highly correlated with the horizontal MDS axis separating Control and Macroalgae meiofaunal communities. Meiofaunal abundance and taxonomic composition in the Bay of Cádiz seem to be bottom-up controlled either through a grazer system based on microphytobenthos in bare sediments or through a decomposer system in macroalgae affected sediments.

Effect of early introduction of microencapsulated diet to larval Atlantic halibut, Hippoglossus hippoglossus L. assessed by microarray analysis.

An experimental microdiet prepared using an internal gelation method was used to partially replace the traditional live feed (Artemia) for larval Atlantic halibut, Hippoglossus hippoglossus L. Three trials were conducted with microdiet introduced at 20, 32, and 43 days post first feeding and larvae were sampled at approximately 2, 13, 23, and 33 days after microdiet introduction in each trial. The success of feeding was assessed by morphometrics and histological analysis of gut contents. Microdiet particles were readily consumed after a period of adaptation and provided an adequate source of nutrients with no significant increase in mortality in the microdiet-fed group compared to the control group. However, growth was limited and there was an increased incidence of malpigmentation of the eye and skin. Subtle changes in underlying digestive and developmental physiology were revealed by microarray analysis of RNA from control and experimental fish given microdiet from day 20 post first feeding. Fifty-eight genes were differentially expressed over the four sampling times in the course of the trial and the 28 genes with annotated functions fell into five major categories: metabolism and biosynthesis, cell division and proliferation, protein trafficking, cell structure, and stress. Interestingly, several of these genes were involved in pigmentation and eye development, in agreement with the phenotypic abnormalities seen in the larvae.

Larval organogenesis of Pagrus pagrus L., 1758 with special attention to the digestive system development.

Organogenesis of the red porgy (Pagrus pagrus L., 1758) was examined from hatching until 63 days post-hatching (dph) using histological and histochemical techniques. At hatching, the heart appeared as a tubular structure which progressively developed into four differentiated regions at 2 dph: bulbus arteriosus, atrium, ventricle and sinus venosus. First ventricle and atrium trabeculae were appreciated at 6 and 26 dph, respectively. Primordial gill arches were evident at 2 dph. Primordial filaments and first lamellae were observed at 6 and 15 dph, respectively. At mouth opening (3dph), larvae exhausted their yolk-sac reserves. The pancreatic zymogen granules appeared at 6 dph. Glycogen granules, proteins and neutral lipids (vacuoles in paraffin sections) were detected in the cytoplasm of the hepatocytes from 4-6 dph. Hepatic sinusoids could be observed from 9 dph. Pharyngeal and buccal teeth were observed at 9 and 15 dph, respectively. Oesophageal goblet cells appeared around 6 dph, containing neutral and acid mucosubstances. An incipient stomach could be distinguished at 2 dph. The first signs of gastric gland development were detected at 26 dph, increasing in number and size by 35-40 dph. Gastric glands were concentrated in the cardiac stomach region and presented a high content of protein rich in tyrosine, arginine and tryptophan. The intestinal mucous cells appeared at 15 dph and contained neutral and acid glycoconjugates, the carboxylated mucins being more abundant than the sulphated ones. Acidophilic supranuclear inclusions in the intestinal cells of the posterior intestine, related to pynocitosis of proteins, were observed at 4-6 dph.

Characterization of a partial alpha-amylase clone from red porgy (Pagrus pagrus): expression during larval development.

A partial alpha-amylase cDNA was isolated from red porgy (Pagrus pagrus, Teleostei: Sparidae) and its tissue specific expression during larval development was examined. The cDNA was 949 bp long and showed 90% identity with other fish amylases. A 545 bp fragment was used to study amylase expression using in situ hybridization and RT-PCR techniques. Both methods showed a similar pattern: high and relatively constant expression for the first 30 days after hatching (dah), subsequently decreasing until the end of the experiment at 60 dah. The goal of this work was to extend the existing knowledge of the functionality of larval fish digestive systems and to provide new information about alpha-amylase gene expression.

Organogenesis of the digestive tract in the white seabream, Diplodus sargus. Histological and histochemical approaches.

The ontogeny of the digestive tract of the white seabream, Diplodus sargus during the larval development up to day 45 post-hatching (dph) has been studied using histological and histochemical techniques. The oesophageal goblet cells appeared around 6 dph and contained neutral and acid mucosubstances (PAS/diastase-PAS and Alcian Blue pH 2.5 positive reactions). An incipient stomach can be distinguished from 2 dph but the first sign of gastric gland development was detected around 13-15 dph, increasing in number and size by 22-23 dph. Gastric glands were concentrated in the cardiac stomach region and they had a high content of protein rich in tyrosine, arginine and tryptophan. Acidophilic supranuclear inclusions related to pynocitosis of proteins, were already observed in the intestinal cells of the posterior intestine around 4-6 dph (exogenous feeding) and they were present until 25 dph. The intestinal mucous cells appeared between 15-18 dph and contained a mixture of neutral and acid mucosubstances/glycoconjugates, carboxylated ones being more abundant than the sulphated ones. The stomach and gastric glands were fully developed by the first month of life marking the beginning of digestive features characteristic of the juvenile stage. Around 4-6 dph, glycogen, proteins and neutral lipids were observed in the granular cytoplasm of hepatocytes. Strongly acidophilic zymogen granules were also present, at this time, in the basophilic cytoplasm of the exocrine pancreatic acinar cells and contained abundant proteins, especially rich in arginine, tyrosine and tryptophan.

Comparative energetics during early development of two marine fish species, Solea senegalensis (Kaup) and Sparus aurata (L.).

Growth, energy content, ingestion and respiration rates and energetic efficiencies were measured in the larvae of two marine fish (Solea senegalensis and Sparus aurata) whose eggs have a similar diameter (approximately 1mm) and energy content (approximately 1J), but whose larvae reach a quite different mass after the first month of life. Experiments were carried out in populations reared under the same conditions in the laboratory during the first month after hatching. Solea senegalensis grow from hatching to the start of metamorphosis (approximately day 14) at twice the rate of Sparus aurata (specific growth rate for Solea senegalensis 0.25 microg day(-1); specific growth rate for Sparus aurata 0.12 microg day(-1)). The tissues in Solea senegalensis larvae accumulated energetic reserves that were used during metamorphosis, which occurred during the third week after hatching. Ingestion and respiration rates differed in the two species during the experimental period. Although Solea senegalensis continued eating during metamorphosis, the specific ingestion rates decreased during the process. Nevertheless, no great differences in specific ingestion rates and rates of oxygen consumption were observed when comparing the same larval mass range. Larvae of both species showed an allometric relationship between respiration rate and biomass. The energetic efficiencies calculated in the present study denoted different metabolic patterns in each species. In Solea senegalensis, the energy used for growth increased progressively during the larval (pelagic) period and then, from the first signs of metamorphic transformation, remained almost constant. In this species, the energy allocated to growth was greater than that allocated to metabolic processes. Sparus aurata invested less energy in growth than in metabolic processes and displayed a constant energy allocation throughout the experimental period. During the first month after hatching, Solea senegalensis always allocated more energy for growth than did Sparus aurata.