Seafood is the number one source of essential fatty acids, particularly, long-chain polyunsaturated fatty acids (LC-PUFA) in the human diet. As global population growth eventually surpasses what the world's wild commercial stocks can provide, reliance on the aquaculture industry to expand production will continue to increase in order to meet the demands of consumers worldwide. Currently, fluctuations in supply and cost coupled with environmental sustainability and contaminant concerns have motivated the aquaculture industry to research alternative lipid sources and feeding strategies in order to reduce the reliance on marine-derived resources. For most cultured species, replacing fish oil with terrestrial plant-based lipid sources is a minor dietary modification that has little consequence on production performance. However, fish raised on these plant-based lipid alternatives contain considerably higher medium chain polyunsaturated fatty acids (MC-PUFA) and n-6 fatty acids and less beneficial LC-PUFA and n-3 fatty acids within the fillets, thus negatively impacting the nutritional value of cultured seafood to the consumer. In order to alleviate this problem, producers can employ finishing strategies to restore fillet LC-PUFA content prior to harvest. As a complement to this approach, provision of dietary saturated fatty acids (SFA) and/or monounsaturated fatty acids (MUFA) in lieu of MC-PUFA appears to maximize the retention of LC-PUFA deposition during the grow-out period and may increase deposition during finishing. Accordingly, my objectives were to 1) assess whether the SFA, MUFA, and MC-PUFA content of the alternative lipid affected LC-PUFA levels in Nile Tilapia fed reduced fish oil feeds; and 2) using the optimal alternative lipid identified in the first objective, assess increasing fish oil replacement rates in conjunction with finishing to maximize product nutritional value and minimize fish oil usage in Nile Tilapia culture. To address the first objective, I assessed production performance and tissue composition of Nile Tilapia fed diets containing fish oil or blends of fish oil and various soybean-derived alternative lipids. Quadruplicate tanks of juvenile Nile Tilapia were fed diets containing fish oil (FISH, high in LC-PUFA) or a 50:50 blend of fish oil and standard (STD-SO, high in MC-PUFA), saturated fatty acid-enriched (SFA-SO, high in SFA), low α-linolenic (LO-ALA-SO, high in MC-PUFA), or hydrogenated (HYD-SO, high in MUFA) soybean oil for 16 weeks. Partial replacement of fish oil with soybean oils did not significantly affect production performance with the exception of the HYD-SO diet which yielded significantly reduced growth efficiency in comparison with some of the experimental diets, though not the FISH control. Despite distinctly different dietary fatty acid profiles, fillet fatty acid composition was similar among fish fed the FISH, SFA-SO, and HYD-SO diets. However, feeding the STD-SO and LO-ALA-SO diets resulted in significant enrichment of less desirable MC-PUFA and n-6 fatty acids within the fillet. Fillet LC-PUFA levels were equivalent among all groups despite the 50% reduction in dietary LC-PUFA intake among fish fed the soybean oil-based feeds. Based on these results, incorporation of STD-SO, SFA-SO, or LO-ALA-SO could be used as partial replacements for fish oil in Nile Tilapia feeds without impairing production performance, though SFA-rich soybean oils appeared to be the best alternative for maintaining a more "fish oil"-associated fillet fatty acid profile. Accordingly, the SFA-enriched soybean oil was selected for further study in the second objective trial that evaluated the effects of graded levels of fish oil replacement without or without implementation of finishing periods on production performance and fillet fatty acid composition. Nile Tilapia were fed feeds containing 100% fish oil (100-FO), the previously assessed SFA-enriched soybean oil (100-SFA-SO), or blends of fish oil and SFA-enriched soybean oil (50-SFA-SO, 75-SFA-SO). Triplicate groups of fish were fed the aforementioned diets exclusively throughout the feeding trial (100-SFA-SO unfinished, 75-SFA-SO unfinished, 50-SFA-SO unfinished) or in conjunction with 4 or 8 weeks of finishing with the 100-FO feed (100-SFA-SO + 4 wks, 100-SFA-SO + 8 wks, 75-SFA-SO + 4 wks, 75-SFA-SO + 8 wks, 50-SFA-SO + 4 wks, 50-SFA-SO + 8 wks) for a total of 20 weeks. Production performance was unaffected by dietary inclusion of SFA-enriched soybean oil when fed exclusively or in combination with fish oil, though growth performance was lower than observed in the previous trial and likely confounded by behavioral interactions and frequent spawning. After 12 weeks of consuming the SFA-enriched soybean oil grow-out diets, fillet levels of n-3 LC-PUFA were not statistically different from 100-FO control levels despite different levels of dietary inclusion. However, the high dietary levels of SFA in the experimental feeds did not translate into increased fillet SFA content, suggesting selective retention of LC-PUFA at the expense of fillet SFA. Finishing for 4 or 8 weeks increased fillet n-3 LC-PUFA content in all groups, though it appears that the 50- and 75-SFA-SO diets were more successful in maintaining acceptable health promoting n-3:n-6 ratios. Based on these results, SFA-enriched soybean oil-based feeds can be used as a cost-saving measure during grow-out, and the effects of these feeds on fillet fatty acid profile can be reversed to a considerable extent in as little as 4 weeks by implementing a finishing period prior to harvest. This approach is a promising strategy for minimizing fish oil usage while maximizing product value of cultured Nile Tilapia.

During preliminary research on cobia (Rachycentron canadum, L.) it became increasingly clear that more in-depth information was required to provide enabling techniques for the cobia aquaculture industry to develop more rapidly. A unifying theme in many of the more important issues facing cobia aquaculture is nutrition. This led to nutritional investigations with larval and juvenile fish highlighting the impacts of dietary ingredients on animal performance. Indeed, nutrition can be viewed as a central lever of action through which many aspects of the physiology and the environmental (water) quality of the animal can be controlled. The first project focused on studying the larval development of cobia, a fish species highly suitable for aquaculture for which the industry is nascent. I described the time-course of development of external sensory organs, gut morphology and relevant digestive enzymes under controlled conditions using electron microscopy, histology and spectrophotometric assays. The developmental sequence of larval cobia could be separated in two phases, with a transition period between 12 and 14 days post hatch (dph). This transition is characterized by the formation of the intestinal loop, the establishment of basic cranial neuromast configuration, leading to the initiation of the onset of pancreatic enzymes and the increase of growth rate. In addition, the effects of dietary taurine supplementation and incorporation of mannan oligosaccharides (MOS) into live feeds on cobia larvae development was examined. Fish fed supplementary MOS did not grow faster but displayed higher microvilli length and density. In addition, MOS-fed fish were more resistant to salinity stress. The dietary supplementation of taurine resulted in a dramatic increase in survival, growth and development rates, and enzymatic activities. The second project aimed at refining cobia juvenile nutrition, assessing fish meal and fish oil replacements. Novel sources, including soy protein and oil, were investigated with and without amino acid and MOS supplementations, yielding promising results. Indeed, both fish meal and fish oil were replaced completely and successfully in feeds for juvenile cobia. In addition, novel ingredients (e.g. marine algae meals and soy protein concentrate) were identified to effectively achieve such replacement. The third and last project dealt with nutrient-gene interactions, specifically centering attention on immunostimulants for which the underlying mechanisms of action remain poorly characterized. Here, dietary MOS, nucleotides and selenomethionine (Se-met) were offered to zebrafish whose transcriptome was analyzed by microarray. The immune system, humoral or cellular, innate or adaptive, exhibited different patterns of response according to the immunostimulating nutrient used. In addition, various genes involved in cell cycle and cytokinesis were concomitantly expressed. An intriguing observation related to the insulinomimetic effect of Se-met. In other words, Se-met impacted pathways normally regulated by insulin, such as the MAPK and PI3K pathways. Some Insulin-like Growth Factors (IGF) and IGF bindgin proteins were up-regulated. Additional research is however necessary prior to advocating for the use of these additives, in order to further investigate their respective pros and cons. / Ph. D.

A whole-cell DHA-rich algal product (A-DHA, provided by Evonik Industries) that is rich in DHA (125 mg DHA/g dry matter) is a possible replacement for fish oil in salmonid diets. The nutrient digestibilities of the algal product were measured in rainbow trout in freshwater and in Atlantic salmon in saltwater (32-33 ppm). In experiment 1, rainbow trout (initial weight ~ 300g) were randomly assigned to 12 x 120 L tanks (n = 10 per tank). A reference diet containing 1% Celite as an indigestible marker and three test diets with increasing percentage of A-DHA substitution (6.67%, 13.33% and 20%) were fed. Feces were collected using a settling column and feed and feces analyzed for digestible dry matter (DM), gross energy (GE), ash, crude protein (CP), essential amino acids and total lipid. The digestibility of six long-chain fatty acids including 18:1n-9 (OA), 18:2n-6 (LA), 18:3n-3 (ALA), 20:4n-6 (ARA), 20:5n-3 (EPA) and 22:6n-3 (DHA) was measured. In experiment 2, Atlantic salmon (~170g) were randomly distributed to 12 fiberglass tanks (600L) with 106 fish per tank. The fish were assigned to four diets with the same levels of A-DHA inclusion as for rainbow trout and yttrium oxide (Y2O3) was used as an inert marker. Feces were collected by stripping and the digestibilities of DM, CP and lipid as well as OA, LA, ALA, ARA, EPA and DHA were determined. In experiment 1, the apparent digestibility of dietary DM, GE and lipid in rainbow trout declined significantly with increasing inclusion of A-DHA (P < 0.01). The inclusion of A-DHA had no effect on the digestibility of CP and ash as well as the availability of essential amino acids (P > 0.05). Furthermore, increased inclusion of A-DHA resulted in significantly lower digestibility of ARA, EPA and DHA (P < 0.05). A similar pattern was seen in the digestibility of OA, LA and ALA, although the effect of A-DHA inclusion was not statistically significant. Regression analysis revealed that nutrient contribution from A-DHA had significantly negative linear and quadratic effects on the apparent digestibility of DM, GE, and lipid. The inclusion levels of A-DHA had both significantly negative linear and quadratic effects on digestibility of LA and ALA, whereas only significantly negative linear effect was found on OA. Significantly negative linear and quadratic regressions were observed for the digestibility of ARA, EPA and DHA. The linear regression for CP was significantly negative and the regressions for the individual amino acids were not significant (P > 0.05). In experiment 2, dietary inclusion of A-DHA had a significantly negative effect on lipid digestibility in Atlantic salmon, at all inclusion rates whereas the significant negative effect on digestibilities of DM and CP was only observed in fish fed 20% A-DHA. The digestibilities of OA, LA, ALA and EPA were greater than 91%. In contrast, the apparent digestibilities of ARA and DHA decreased significantly with increasing substitution of A-DHA (P < 0.01). Significantly negative linear and quadratic regressions were found between nutrient contribution from A-DHA to the diets and apparent digestibility of DM, CP and lipid, so were LA, EPA and DHA. However, there were only significant quadratic regressions for OA, ALA and ARA, but not significant linear effects. Subsequently, a twelve-week feeding trial in rainbow trout was conducted to investigate the impact of replacing fish oil with A-DHA in canola-oil-based diets on the growth performance and fatty acid composition and retention. Four experimental diets containing only canola oil (CO; 13.5%), fish oil (FO; 13.5%), canola oil and fish oil (C+F; 7.4% and 6.1%, respectively) or canola oil and A-DHA (C+A; 15.5% and 6%, respectively) were formulated to contain 386.2 g/kg digestible crude protein and 17.58 MJ/kg digestible energy. In addition, the C+A diet was formulated to have the same DHA concentration as in the C+F diet. Each diet was fed to three tanks of rainbow trout (average initial weight of 70g; n = 17/tank) and the fish were fed to apparent satiation 2 times daily. At the end of the growth trial, all fish approximately tripled their weight. No significant differences were noted between the dietary treatments in growth performance as measured by final weight, average weight gain, feed intake, specific growth rate (SGR) and feed conversion ratio (FCR). Although FO and C+A fed fish tended to accumulate more lipids, final whole body lipid content did not differ significantly between dietary treatments (P = 0.11). The concentrations of EPA, DHA as well as total n-3 fatty acid were significantly higher in fish fed the FO diet than fish fed the other 3 diets. The C+A fed fish had lower EPA and higher DHA concentrations compared with the CO and C+F fed fish; however, the differences were not significant. Apparent retention of total lipid in the trout was not significantly influenced by treatments (P > 0.05). Similarly, dietary treatments had no significant effect on the apparent retention of total saturated fatty acids, total mono-unsaturated fatty acids, n-3 polyunsaturated fatty acids and n-6 polyunsaturated fatty acids. The retention of 18:4n-3 (SDA) was significantly higher (> 100%) in fish fed CO and C+A compared with fish fed FO and C+F (< 51%), indicating greater bioconversion of ALA to SDA in the CO and C+A fed fish than in FO and C+F fed fish. The retention of EPA in the CO and C+A fed fish was over 100%, suggesting a net synthesis of EPA in these treatment groups. In contrast, the EPA retention in the FO and C+F fed fish was 55 and 21%, respectively, which showed a tendency to be significantly lower than that in the other two groups (P = 0.09). The CO fed fish had significantly higher DHA retention than fish fed the other 3 diets. The DHA retention in the FO fed fish (112%) was numerically but not significantly higher than in the C+F (66%) and C+A fed fish (73%). Thus, feeding the C+A to rainbow trout resulted in DHA retention equal to feeding the C+F.