Silvia Martínez-Llorens

Impact of Fishmeal Replacement in Diets for Gilthead Sea Bream (Sparus aurata) on the Gastrointestinal Microbiota Determined by Pyrosequencing the 16S rRNA Gene

Recent studies have demonstrated the impact of diet on microbiota composition, but the essential need for the optimization of production rates and costs forces farms and aquaculture production to carry out continuous dietary tests. In order to understand the effect of total fishmeal replacement by vegetable-based feed in the sea bream (Sparus aurata), the microbial composition of the stomach, foregut, midgut and hindgut was analysed using high-throughput 16S rDNA sequencing, also considering parameters of growth, survival and nutrient utilisation indices.A total of 91,539 16S rRNA filtered-sequences were analysed, with an average number of 3661.56 taxonomically assigned, high-quality sequences per sample. The dominant phyla throughout the whole gastrointestinal tract were Actinobacteria, Protebacteria and Firmicutes. A lower diversity in the stomach in comparison to the other intestinal sections was observed. The microbial composition of the Recirculating Aquaculture System was totally different to that of the sea bream gastrointestinal tract. Total fishmeal replacement had an important impact on microbial profiles but not on diversity. Streptococcus (p-value: 0.043) and Photobacterium (p-value: 0.025) were highly represented in fish fed with fishmeal and vegetable-meal diets, respectively. In the stomach samples with the vegetable diet, reads of chloroplasts and mitochondria from vegetable dietary ingredients were rather abundant. Principal Coordinate Analysis showed a clear differentiation between diets in the microbiota present in the gut, supporting the presence of specific bacterial consortia associated with the diet.Although differences in growth and nutritive parameters were not observed, a negative effect of the vegetable diet on the survival rate was determined. Further studies are required to shed more light on the relationship between the immune system and sea bream gastrointestinal tract microbiota and should consider the modulation of the microbiota to improve the survival rate and nutritive efficacy when using plant-based diets.

Effect of dietary protein level and source on digestive proteolytic enzyme activity in juvenile Senegalese sole, Solea senegalensis Kaup 1850

The effect of dietary protein level and protein source on growth and proteolytic activity of juvenile Solea senegalensis was studied. In Experiment 1, fish were fed on four experimental diets containing increased protein levels (36, 46, 56 and 67%). In Experiment 2, Senegalese soles were fed on five diets with partial substitution of fish meal by soybean meal, soybean protein concentrate, soybean protein isolate, wheat gluten meal or pea protein concentrate. Results prove that growth and proteolytic activity in the distal intestine of fish were affected by the quantitative increase in dietary protein. The origin of protein source used in the elaboration of experimental diets affected both the amount and composition of the alkaline proteases secreted into the intestinal lumen; however, it did not decrease animal growth. Juvenile Senegalese sole showed capability to modulate digestive protease secretion when the concentration and/or source of dietary protein were modified. Quantity and quality of dietary protein affected protein hydrolysis in Senegalese sole intestine. This study establishes that 30% fish meal protein can be replaced by soybean derivatives without affecting intestinal proteases. Replacement with wheat gluten meal or pea protein concentrate should be taken cautiously, but further research is needed to establish whether growth performance and digestive enzyme physiology of Senegalese sole are affected by plant protein-supplemented diets in a long-term trial.

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

ABSTRACT 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.

The effect of dietary soybean meal on growth, nutrient utilization efficiency, and digestibility of juvenile common dentex, Dentex dentex (Actinopterygii: Perciformes: Sparidae).

Background. The aim of this work was to study the growth of juvenile dentex fed on cooking-extruded diets, determine the level of substitution of fishmeal by soybean meal may be without affecting the growth in this species and its digestibility. MaterialsandMethods. The availability of defatted soybean meal as a substitute for fish meal was evaluated in juvenile (41 g on average) dentex by feeding diets containing 0%-60% soybean meal, for 97 days. Results. Survival at the end of the experiment was high (80%) except for the fish fed diets with 50% and 60% substitution. Growth, feed gain ratio, and protein efficiency ratio (PER) were slightly reduced at higher soybean meal levels. Fish fed diets containing 0% to 40% of soybean meal grew significantly more and FCR was lower than fish fed other diets. No differences were obtained for protein digestibility coefficients of experimental diets (20%, 30%, 40%, and 50% SBM). Conclusion. The results confirm the best protein level for optimum growth seems to be around 50% and 12% lipid level, and maximum soybean meal substitution of 40%.

A study of partial replacement of fish meal with sunflower meal on growth, amino acid retention, and body composition of sharpsnout seabream, Diplodus puntazzo (Actinopterygii: Perciformes: Sparidae)

Background. Our previous study demonstrated that sharpsnout seabream,Diploduspuntazzo (Walbaum, 1792) , can be fed with up to 34.8% sunflower meal (SFM), with excellent results in growth parameters and feed effi - ciency. The aim of the current study was to test the replacement of fish meal with SFM in the diet formulation for sharpsnout seabream and to evaluate growth, nutritive parameters, amino acid retention, and body composi - tion of the fish during the fattening period. Materialsandmethods. Sixteen baskets (300-L capacity), each with thirteen fish, were distributed in a recircu - lated saltwater system to allow four experimental diets containing 40% crude protein (CP) and 20% crude lipid (CL) with 0%, 11.7%, 23.5%, and 34.8% SFM partially replacing fish meal to be used in quadruplicate for this experiment. The fish were fed these dietsadlibitum during the experiment, which lasted for 162 days. Growth, nutrition efficiency, biometrics, carcass composition, amino acid composition, and amino acid retention of the experimental fish were evaluated. Results. There were no statistical differences in the growth parameters among the treatments. However, fish fed a diet containing 34.8% SFM had the lowest feed intake (FI), lowest feed conversion ratio (FCR), and the high - est protein efficiency ratio (PER). There were also no statistical differences in the biometric parameters although fish fed the diet containing 10% SFM had the lowest CP levels. Leucine was the only difference in the essential amino acid (EEA) profile with fish fed a diet containing 11.7% SFM having the lowest levels of leucine. Although there were fluctuations among the amino acid retentions, they were not statistically significant. Conclusion. SFM (up to 34.8%) can be included in the diets of sharpsnout seabream, thereby, replacing 27% of the fish meal without altering the fish growth.

Fish oil substitution by soybean oil in Sharpsnout seabream Diplodus puntazzo: Performance, fatty acid profile, and liver histology

ABSTRACT The present study was conducted to determine the effect of soybean oil on the performance and liver histology in Sharpsnout seabream (Diplodus puntazzo) (initial body mass 35 g). Four experimental diets were formulated containing 0%, 25%, 50%, and 75% of soybean oil substituting fish oil. Fish were fed for 84 days. Increasing the level of soybean oil had no significant effects on growth and feed efficiency parameters. Biometrics, body composition, protein efficiency, and energy efficiency were not affected by the fish oil replacement. Muscle and liver fatty acids reflected fish oil substitution. Moreover, histology did not show statistical differences among treatments. Sharpsnout seabream juveniles can be fed with diets in which fish oil has been replaced with soybean oil at up to 75% of total lipid in the diet for a period of 84 days without affecting growth, feed efficiency, biometric parameters, and body composition.

Inclusion of alternative marine by-products in aquafeeds with different levels of plant-based sources for on-growing gilthead sea bream (Sparus aurata, L.): effects on digestibility, amino acid retention, ammonia excretion and enzyme activity

ABSTRACT The search for new sustainable aquafeeds for the species with greater economic importance, such as the gilthead sea bream in Europe, is one of the main challenges in the aquaculture sector. The present work tested fishmeal replacement by a mixture of plant meals at different levels, as well as the use of marine by-products with attractant properties and high-quality protein in high plant protein diets. In order to do that, effects on growth and biometric parameters, digestibility, amino acid retention, excreted ammonia and proteases and amylase activity were assessed, using six different diets: FM100 (100% of protein provided by fishmeal), FM50 (50% of replacement), FM25 (75% of replacement) and FM0 (100% of replacement), but also FM25+ (75% of replacement and 15% of squid and krill meal inclusion), and FM0+ (100% of replacement and 15% of squid and krill meal inclusion). In group FM0, a clear impact of dietary changes was observed on growth, survival and ammonia excretion. Amino acid retention in group FM0+ was also significantly affected, which can be explained by the limited content of certain amino acids in this diet. On the other hand, no significant differences were observed in most biometric parameters or in enzyme activity. In conclusion, complete fishmeal replacement can be achieved by using a mixture of plant-based sources, but supplementation with complementary marine ingredients can prevent detrimental effects on growth, survival, nutritional parameters and protein metabolism.

Pea protein concentrate in diets for sharpsnout sea bream (Diplodus puntazzo): effects on growth and health status

ABSTRACT Four diets for sharpsnout sea bream juveniles (14 g body weight) with four levels of air-processed pea protein concentrate (PPC) (0, 160, 320 and 487 g/kg diet) were tested in triplicate. The experimental diets were isonitrogenous (43% crude protein) and isolipidic (19% ether extract) and the fish were fed to satiation twice a day. After 125 d, fish growth was diminished by the inclusion of PPC. Feed conversion did not show significant differences in any treatment. Neither the body analyses nor the protein and individual essential amino acid retention efficiencies were affected by the inclusion of PPC in the diet. However, histological gut examinations revealed noticeable differences. Fish fed the diet with the highest inclusion level of PPC presented the longest villous length and the most goblet cells, and the width of the lamina propria increased in the anterior intestine. Although no negative changes in nutritive parameters were detected, these alterations might affect nutrient transport, with negative consequences for fish growth. It was concluded that the PPC in the amounts tested here is an inappropriate substitute for fishmeal in diets for sharpsnout sea bream juveniles.