Evaluation of Phytase Impact on In Vitro Protein and Phosphorus Bioaccessibility of Two Lupin Species for Rainbow Trout (Oncorhynchus mykiss).

Legumes are an important source of protein, lipids, and other essential nutrients. As the demand for protein and lipids continues to surge on a global scale, there is a growing interest in incorporating legumes into aquafeeds. This shift is driven not only by the escalating growth of the aquaculture sector in recent years but also by the imperative to diminish the dependency on traditional resources like fishmeal (FM) and fish oil. Amongst legumes, different lupin species had been identified as a potential protein source to partially reduce the inclusion of FM in countries such as Australia, Chile, and the European Union. A comprehensive evaluation of their nutritional profiles, overall characteristics, and potential antinutritional factors is essential for informed utilization and the implementation of nutritional enhancement strategies. In pursuit of this goal, an in vitro gastrointestinal simulation system was devised to replicate the digestive conditions of rainbow trout (Oncorhynchus mykiss). The study focused on determining the bioaccessibility of protein and phosphorus within two sweet lupin varieties (alkaloids < 0.05) with high (Lupinus mutabilis) and low (Lupinus angustifolius) native phytic acid content evaluated as whole (W) or dehulled (D) seeds meals and the effect of a single dose of phytase (2,500 FTU/kg DM). Additionally, regular soybean meal (SBM) served as reference (10 treatments with 3 replicates). A 2,500 FTU/kg DM phytase dose increased the levels of PO4-3 released throughout the intestinal phase by 122.6% for L. mutatabilis W, 116.3% for L. mutatabilis D, 65.2% for L. angustifolius W, 59.0% for L. angustifolius D, and 91.8% for SBM compared to controls without phytase. The bioaccessibility of amino acids in varieties treated with phytase increased with respect to the control without phytase. L. mutabilis was found to be a potentially viable alternative as a good quality protein source for the manufacture of environmentally friendly aquafeeds.

Potential use of high levels of vegetal proteins in diets for market-sized gilthead sea bream (Sparus aurata).

The effect of partial or total dietary substitution of fishmeal (FM) by vegetal protein sources on growth and feed efficiency was carried out in on-growing gilthead sea bream (mean initial weight 131 g). The Control diet (FM 100) contained FM as the primary protein source, while in Diets FM 25 and FM 0 the FM protein was replaced at 75% and 100%, respectively, by a vegetable protein mixture consisting of wheat gluten, soybean meal, rapeseed meal and crystalline amino acids. Diets FM 25 and FM 0 also contained krill meal at 47 g/kg in order to improve palatability. At the end of the trial (after 158 d), fish survival was above 90%. Final weight and the specific growth rate were statistically lower in fish fed the Control diet (361 g and 0.64%/d), compared with 390-396 g and 0.69-0.70%/d after feeding vegetal diets. No significant differences were found regarding feed intake and feed conversion ratio. The digestibility of protein and amino acids (determined with chromium oxide as indicator) was similar in all diets. The blood parameters were not significantly affected by treatments. The activity of trypsin and pepsin was significantly reduced after feeding Diet FM 0. In the distal intestine, the villi length in fish fed Diet FM 25 was significantly longer and the intestine of the fish fed the FM 100 diet showed a smaller number of goblet cells. In conclusion, a total FM substitution by a vegetal mix supplemented with synthetic amino acids in on-growing sea bream is feasible.

In vitro characterization of acid secretion in the gilthead sea bream (Sparus aurata) stomach.

The gastric acid secretion of juvenile Sparus aurata was characterized in Ussing chambers; secretion rates were determined by a pH-stat method at pH5.50 and bioelectrical parameters were measured in current-clamped tissues. The basal secretion equaled to 535±87nmol·cm(-2)·h(-1). Serosal carbachol 100µM produced an increase (ΔJH(+)) of 725±133nmol·cm(-2)·h(-1) from basal secretion, this effect being inhibited by mucosal omeprazole 100µM. Basal secretion was also sensitive to the combination of serosal forskolin (FK) 10µM+serosal isobutylmethylxanthine (IBMX) 100µM (ΔJH(+)=793±239nmol·cm(-2)·h(-1)); this effect was insensitive to mucosal omeprazole 100mM but inhibited by mucosal bafilomycin A1 100nM. The effect of carbachol proceeded within a few minutes (<10min), whereas the effect of FK+IBMX was gradual, taking 40min to reach the maximum. The addition of mucosal gadolinium (Gd(3+)) 100µM, a potent calcium-sensing receptor (CaR) agonist, stimulated the basal secretion (ΔJH(+)=340±81nmol·cm(-2)·h(-1)). The present results indicate that the acid secretion mechanism in the sea bream stomach is regulated by muscarinic and CaR-like receptors, cAMP is implicated in the signal transduction, and at least two proton pumps, a HK-ATPase and a V-ATPase contribute to acid secretion.

Gut pH as a limiting factor for digestive proteolysis in cultured juveniles of the gilthead sea bream (Sparus aurata).

After the development of the gastric function in juvenile fish, dietary proteins enter a two-phase digestive process comprising an acidic gastric phase followed by an alkaline intestinal phase. However, the main gastric protease, pepsin, is strictly dependent on the existence of a low-enough environmental pH. In 20-g gilthead sea bream, Sparus aurata, the mean minimal gastric pH is close to 4.5, while the mean pH in the duodenal portion of the intestine was nearly fixed at 6.5. The mean maximal gastric content of HCl was approximately 20 microEq for a low-buffering diet. Gastric proteases were more severely affected than intestinal proteases when assayed at actual sub-optimal pH values, 4.5 and 6.5, respectively. When the gastric proteases of juvenile fish were pre-incubated with a citric acid buffer at pH 6.0, the activity at pH 4.5 was very low, whereas when they were pre-incubated with the same buffer at pH 3.0, the activity at pH 4.5 was significantly increased; this fact suggests a deficient activation of zymogens during the gastric digestion and points to a potential approach to improve protein digestion in juvenile gilthead sea bream.