Currently aquaculture is the largest growing food sector in the world, however, its future growth is limited by its heavy reliance on fishmeal (FM). Overfishing of wild marine fish stocks used for FM is putting too much pressure on the stocks, making FM unsustainable as a dominant protein source as aquaculture continues to grow. Plant proteins (PP) are an ideal alternative to FM because of their wide availability and relatively low cost. Soybean meal (SBM) is the most commonly used PP in aquaculture, but its inclusion in feeds is limited due to reduced digestibility and the presence of antinutritional factors ultimately leading to a reduced growth performance of fish that are fed with the SBM-based diet. Therefore, the goal of this thesis was to mitigate the negative effects of PP and improve the growth of fish fed with a PP-based diet, utilizing SBM as our PP in feeds. Three feeding trials were conducted to test the efficacy of 3 approaches towards improving the use of PP in fish. The first trial (Chapter 2), tested the effect of nutritional programming (NP) on the utilization of SBM in zebrafish (Danio rerio). NP is the theory that an organism can be ‘programmed’ to better utilize a dietary component by being exposed to that component in its early life stages. This study also tested the effect of NP through the broodstock by exposing the broodstock to SBM for 4 weeks prior to breeding. In addition, a combination of both programming techniques was also tested. The results found that neither of the programming techniques had a significant effect on the growth performance of the zebrafish. Among the two groups from the programmed broodstock, the group that also received early stage NP had a significantly higher expression of PepT1, a di- and tri- peptide transporter. Also, the dual programmed group had the highest length-to-width ratio of the distal villi among groups that were fed SBM, which signifies an increase in surface area for nutrient absorption in the intestine. The findings from this study suggest that early stage NP may increase the absorption of nutrient from PP-based feeds within the intestine. The second feeding trial (Chapter 3), utilized the supplementation of health-promoting dipeptides to improve the use of SBM-based feeds for zebrafish. The three dipeptides used in this study were alanyl-glutamine, carnosine, and anserine. The five groups in this study consisted of three groups receiving an SBM-based diet with one of the dipeptides supplemented into it, a (-) Control group receiving a non-supplemented SBM-based diet, and a (+) Control group receiving a FM-based diet. Both the alanyl-glutamine and carnosine supplemented groups experienced a significantly higher weight gain compared to the (-) Control group. In addition, the alanyl-glutamine supplemented group had a significantly higher length-to-width ratio of the intestinal villi and, had a numerically higher expression of both nutrient absorption genes measured, PepT1 and fabp2, compared to the (-) Control group. This finding suggests that the supplementation of alanyl-glutamine into SBM-based diets may improve the intestinal absorptive capacity of the fish fed with the SBM. The results from this study also support the use of both alanyl-glutamine and carnosine supplementation as a means of improving the growth performance of fish fed with a SBM-based diet. The third feeding trial (Chapter 4), was conducted on largemouth bass (LMB) (Micropterus salmoides). This study also focused on NP as a means of improving the utilization of dietary SBM, similar to Chapter 2. In this study, live feed was used as a vector to program the larval LMB to SBM. The programmed group in this study received Artemia nauplii that were enriched with a SBM solution, during the larval stage. The bass were then fed with a FM-diet for 7 weeks, before being reintroduced to SBM for the final 6 weeks of the study (PP-Challenge). The programmed LMB experienced a significantly higher weight gain compared to the non-programmed fish also undergoing the PP-Challenge, and achieved a weight gain similar to that of the LMB that were being fed with an FM-diet. In addition, the programmed LMB had significantly longer distal villi and a higher length-to-width ratio of the villi, compared to the non-programmed group. The findings from this study support the use of live feed as a vector for NP and improving the growth performance of a carnivorous aquaculture species fed with a SBM-based diet. The overall findings from these studies suggest that both NP and dipeptide supplementation are feasible means of improving the utilization of SBM in fish. The mechanism behind NP seems to lie in the intestine. In both zebrafish and LMB, NP was found to reduce the inflammatory impact on the intestine and increase the surface area for absorption of the intestinal villi. The supplementation of alanyl-glutamine had similar effects on the intestine as NP and improved the growth performance of zebrafish. The observations from these studies seem to point to mitigating the negative effects of SBM on the intestine as the key to improving the growth performance of fish fed with a SBM-based diet.

Submitted by Caroline Falcao (caroline.rfalcao@ufpe.br) on 2017-05-22T17:37:54Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese-Aurelyanna- OCEANOGRAFIA.pdf: 849615 bytes, checksum: 58c710c655e4bb0252be1dd111aa04c0 (MD5) / Made available in DSpace on 2017-05-22T17:37:54Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese-Aurelyanna- OCEANOGRAFIA.pdf: 849615 bytes, checksum: 58c710c655e4bb0252be1dd111aa04c0 (MD5) Previous issue date: 2014-08-22 / O alimento vivo é um fator fundamental para o desenvolvimento larval de crustáceos decápodes. Copépodos se destacam como alimento vivo para larvas carnívoras devido ao seu excelente valor nutricional, alta digestibilidade e tamanho adequado. Porém, o uso destes organismos em larviculturas ainda depende do aprimoramento das técnicas de cultivo. O presente trabalho foi dividido em três capítulos. O objetivo dos capítulos 1 e 2 foi aprimorar a técnica de cultivo do copépodo harpacticóide Tisbe biminiensis e testar novas dietas. O cultivo dos copépodos foi realizado em volume de 15L em bandejas plásticas de 0,37m2 adaptadas com drenos para facilitar a troca de água e coleta da prole. No Capítulo 1 foram testadas as rações Alcon Basic® Dieta Controle (RC), ração experimental para peixes Op0 (RE), ração comercial para peixes Nutripeixe AL55® (RP) e ração comercial para camarão marinho Camaronina CR1® (RCM). Apesar da boa aceitabilidade de todas as rações, diferiram de RC a ração RE, apresentando valores inferiores de produção de prole do início dos experimentos até o primeiro pico de produção e a ração RCM, que apresentou resultados inferiores na fase de estabilização da população total e no pico de produção de prole. A produção diária por caixa de cultivo foi de 9.000 ind. L-1 ou 364.864 ind m2 . No Capítulo 2 a dieta controle (RC) foi comparada a dieta teste (RT) ração comercial para peixes marinhos, NRD 5/8, INVE. Durante o período experimental, a produção de prole obtida com a RT não diferiu da produção obtida com a RC, sendo em média 7.400 ind. L-1 . dia -1 ou 300.000 ind. m2 . No Capítulo 3 foi avaliado o desempenho da prole de T. biminiensis como substituta dos náuplios recém-nascidos de Artemia, na alimentação das pós- larvas (PL1 a PL10) do camarão marinho Litopenaeus vannamei. Três tratamentos foram testados: T1 – Controle Artemia, T2 – Mix: Artemia + T. biminiensis e T3 – T. biminiensis. A sobrevivência no T1 foi significativamente maior em comparação com as demais (T1 100% > T2 52% = T3 51%). O comprimento larval foi significativamente maior no tratamento T2 (T2 7.9 mm > T1 7.1 mm = T3 6.8 mm) e não houve diferenças no peso seco. As adaptações realizadas nos sistemas de cultivo reduziram o esforço de coleta e a boa aceitação de T. biminiensis aos diversos tipos de ração reduziram os custos de produção. A quantidade produzida ainda é baixa, mas pode satisfazer a necessidade de setores como a aquicultura ornamental. Os bons resultados alcançados em termos de crescimento e peso no T2 indicam que a combinação de alimentos melhora a qualidade das pós-larvas. A menor sobrevivência das pós-larvas de L. vannamei nos tratamentos T2 e T3 podem estar relacionada a predação, pelos copépodos, de pós-larvas debilitadas. A presença de bactérias patogênicas provindas dos cultivos de copépodos deve ser investigada no futuro. Os resultados indicam que na ausência ou restrição de cistos de Artemia no mercado o cultivo de copépodos poderia vir a representar uma alternativa para as larviculturas desta espécie. / The live food is essential for optimal larval development of decapod crustaceans. Copepods offer excellent nutritional quality, a high degree of digestibility and an adequately small size. However, the use of these organisms in hatcheries still depends on the improvement of farming techniques. This work was divided into three chapters. The aim of Chapters 1 and 2 was to improve the technique of cultivation of harpacticoida copepod Tisbe biminiensis and test new diets. The cultures were performed in plastic trays (0.37 m2 ) in volumes of 15 L with drains to facilitate the water exchange and collection of the offspring. In chapter 1, were tested control diet Alcon Basic® (CD) an experimental diet (ED) for Rachycentron canadum juveniles Op0; a commercial diet Nutripeixe AL55® (FD) and a commercial diet for marine shrimp Camaronina CR1® (MSD). In comparison to the CD treatment, the ED led to significantly lower offspring production from the beginning of the experiment through to peak production and the MSD achieved significantly poorer results regarding the stabilization phase of the overall population and peak offspring production. The results of the daily production per cultivation unit was 9000 ind. L-1 or 364,864 ind m2 . In Chapter 2 the control diet (CD) was compared to test diet (TD) commercial feed for marine fish, NRD 5/8, INVE. During the experimental period, the production of offspring obtained with TD did not differ in yield with the RC, averaging 7,400 ind. L-1 . ind day -1 or 300,000. m2 . In Chapter 3 was evaluated the performance of offspring of the T. biminiensis like substitute of nauplii of Artemia in feeding post-larvae (PL1 to PL10) of the marine shrimp Litopenaeus vannamei. Three treatments were tested: T1 - Artemia Control, T2 - Mix: T. biminiensis + Artemia and T3 - T. biminiensis. The Survival in T1 was significantly higher compared with the other treatments (T1 100% > T2 52% = T3 51%). Larval length was significantly higher in T2 (T2 7.9 mm > T1 7.1 mm = T3 6.8 mm) and no differences in dry weight. The adaptations carried in plastic trays reduced the harvest effort and the. The good acceptance of T. biminiensis the different types of diet reduce production costs. The number of offspring produced is still low, but it can satisfy the sectors such as ornamental aquaculture. The good results achieved in T2 (growth and weight) indicate that the combination of foods improves the quality of post- larvae. The lower survival of post-larvae in treatments T2 and T3 may be related to predation of the weakened post-larvae by copepods. The presence of pathogenic bacteria in cultures of copepods should be investigated in the future. The results indicate that in the absence or restriction of Artemia cysts, the copepods cultures may represent an alternative to the hatcheries.