A feeding experiment was designed to examine how fish growth rates are affected by the size abundance of pelagic zooplankton. Juvenile chinook salmon, Oncorhynchus tshawytscha , were raised in 90 m³ mesh enclosures in Saanich Inlet, B.C. The enclosures permitted ample water and zooplankton circulation while retaining 5-6 gram juvenile salmon. The enclosed fish grew at an average rate of 1.8% wet body weight/day for a six week experimental period. Weekly growth rates ranged from 3.9%/day while food was abundant, to -0.5%/day when food was scarce. Several analytical methods were used to establish a relationship between fish gr.owth and the size and abundance of zooplankton in the enclosures. There is a strong relationship between the fish growth rates and the abundance of 1.4-4.5mm prey. Rates of successful search vary directly with the size and inherent contrast of a prey item. A minimum rate of successful search of 2.0m³/hour was estimated from a functional response curve for salmon feeding on 1.4-4.5mm zooplankton. This value is discussed in relation to a salmonids physical capabilities and results from previous field studies and tank experiments. Daily growth increments on the otoliths of the enclosed fish were examined with respect to daily variations in water temperature and zooplankton abundance. Extremes in food abundance appear to have a significant and consistent effect on the spacing of the growth rings. However, water temperatures would have to be kept constant in order to establish any closer relationship between food abundance and otolith growth rings. / Science, Faculty of / Zoology, Department of / Graduate

Previous studies have demonstrated that the feeding behaviour of Lumbriculus variegatus may be significantly inhibited during exposure to toxic substances. The potential use of an in situ sediment bioassay, using L.variegatus post-exposure feeding inhibition as an endpoint, was investigated. The bioassay consisted of exposing animals in the field for a six-day exposure period and feeding rates were measured immediately afterwards over a twenty-four hour post-exposure period. The bioassay methodology developed in the laboratory produced a consistent baseline response that was reliable and repeatable. Endpoint sensitivity was demonstrated under laboratory conditions, where bioassay organisms exhibited delayed recovery from feeding inhibition after previous exposure to sediment-associated contaminants. The apparent insensitivity of the bioassay to sediment-associated metals means that the technique should only be used as part of a suite of bioassays that employ representative deposit feeders. The ecological relevance of the bioassay endpoint was also demonstrated by comparing short-term measures of post-exposure feeding inhibition with the longer-term effects of a toxicant on L.variegatus populations. The bioassay methodology was successfully adapted for in situ use. Post-exposure feeding inhibition was detected at contaminated field sites. However, the consistent baseline response produced in the laboratory could not be replicated during deployments of the bioassay at upstream (“clean”) field sites. Increased environmental “noise” may have been a result of a number of confounding factors that could limit the sensitivity of the bioassay endpoint if not adequately controlled. Despite the above concerns, the in situ bioassay is suggested to represent a useful tool, which uses a more realistic field exposure scenario to investigate the effects of sediment-associated toxicants with an important functional component of aquatic ecosystems.

Fish meals (FM) and fish oils (FO) are used extensively as the main protein and lipid sources respectively in industrially compounded salmon feeds, mainly due to their excellent nutritional properties. Nevertheless, several reasons dictate the utilisation of sustainable alternative protein and lipid sources and the subsequent reduction in the reliance on fishery-products in aquafeeds. Soy protein concentrate (SPC) is a very promising alternative to FM Hence, the main objective of the present thesis was to investigate the effects of the increased substitution of FM withSPC, lysine and methionine on the growth performance and immune responses of early and late stage Atlantic salmon parr prior to and after vaccination with commercial vaccines (Experiments II and V presented in Chapters 4 and 6). Furthermore the effects of increasing dietary levels of SPC with consistent and/or increasing dietary supplementation of phosphorus on the Atlantic salmon parr’ body proximate and mineral composition or the skeletal mineral composition respectively, prior and post-vaccination with commercial vaccines; under continuous light or under 12 hours light: 12 hours dark photoperiod (Experiments I and V presented in Chapters 3 and 5 respectively). Lastly the impact of FM-, FO- and fishery-free diets on the growth, carcass proximate composition and immune status of Atlantic salmon post-smolts was investigated (Experiment III presented in Chapter 7). In the latter experiment six diets were tested including: two commercially applied marine based diets, one with partial inclusion of vegetable proteins (VPs) and oils (VOs) according to the EU standards (2011-12) (MBE) and one with partial inclusion of VPs, VOs and land animal-by product (ABP) proteins according to the non-EU standards (MBABP); a fully vegetable protein (VP) diet; a fully algal and vegetable oil (VO) diet; a marine-free VP and VO and algal oil diet (VP/VO) diet; and a marine-free diet with a mix of VPs and land ABP proteins and lipid from VOs and algal oils (MFABP). The results of the Experiments I and II (Chapters 3 and 4) show that late Atlantic salmon parr can grow efficiently on SPC80 diets, however, they require longer periods to adapt to these diets compared to fish fed diets containing lower levels of SPC as a protein source. Decreased mineralisation of body cross-section was observed for salmon fed increasing dietary SPC. Vaccination improved mineralisation for the high dietary SPC salmon groups. However, continuous light exposure promoting fast growth appeared to be detrimental for Atlantic salmon body cross ash, Ca, Mg, Mn, P and Zn. Moreover, it was demonstrated that substitution of up to 50% of high quality FM protein with SPC and constantly added P has the minimum possible impact on late salmon parr growth, whereas it stimulates several immune parameters prior to immunisation. Immunostimulatory effects were also shown for the diets with higher dietary SPC levels. It is not clear if these results were an effect of increased FM replacement with SPC or not properly balanced levels of dietary P. The Experiments III and IV (Chapters 5 and 6) illustrated that early stage Atlantic salmon parr can accept diets with up to 58% protein from SPC without serious effects on body growth. However, higher levels can severely affect salmon growth performance. Moreover, it was shown that long-term feeding of salmon with increasing dietary SPC combined with increasing phosphate supplementation, alone or in combination with vaccination can actually be beneficial for Atlantic salmon parr bone mineralisation. However, mineralisation in vaccinated fish was higher than in PBS-injected fish. This could be linked to the slower growth of vaccinated salmon allowing their developing bones to mineralise properly. Changes in the modulation of the different components of the complement activity was revealed in Atlantic salmon fed increasing dietary levels of SPC. The modulation of complement activity was demonstrated at both studies utilising increasing dietary SPC concentrations, indicating that complement componenets are among the most prominent immunological markers upon dietary FM replacement with SPC. However, overall no differences in total complement activity and therefore the immune capacity and resistance against Aeromonas salmonicida were observed among the salmon groups receiving increasing levels of SPC. Lastly in Experiment V (Chapter 7) higher growth performance indices (weight gain, SGR and TGC) were evident in the MBE salmon compared to the MBABP group, salmon fed diets with complete eleimintion of FM or FO (VP and VO respectively) and fish fed two diets with total substitution of both marine derived feed ingredients, three months after the start of the feeding trial. Higher feed intake was demonstrated for both MBE and VP salmon compared to the other groups for the duration of the first period. The above results could have been influenced by discrepancies in the size of the fish at the start ofthe trial, revealing flaws in the experimental design. Both MBE and VP salmon groups also presented the highest feed conversion ratios, revealing the lowest efficiency in dietary nutrient utilisation in comparison to the rest of salmon which exhibited no differences in feed efficiency, revealing an overall better performance of the MBABP and diets with low levels of marine feedstuffs. Improved FI compared to the values of the first period and higher SGR and TGC values were demonstrated for salmon from the latter treatments compared to MBE and VP salmon, during the second part of the study, revealing compensatory growth for these groups. VP salmon demonstrated the highest and VO salmon the lowest condition factor values. The former finding might possibly be related with higher fat accumulation in the viscero-hepatic tissues. No differences were observed in carcass moisture, protein, fat and ash concentrations among the dietary groups of fish. Furthermore, no differences were demonstrated in terms of total and differential leucocyte counts, plasma haemolytic activity, plasma protein and total IgM levels, stimulated and non-stimulated HKM burst activity among the different dietary groups. However, lower haematocrit values were observed in the MB and VO-fed groups compared to the MFABP and VP/VO groups. Furthermore, decreased lysozyme activity was observed for all diets in contrast to the control groups, whereas FM-free diets promoted plasma anti-protease activity. The former result could have been an effect of either immune or stress induction, whereas the second is regarded as an immunostimulatory effect. The results suggest that marine-oil, marine-protein and marine-free diets could be satisfactorily used for Atlantic salmon post-smolts without severe reductions in their innate immune responses, although longer adaptation periods might be required for the fish to fully accept these diets.

The current general economic and food crises have generated an unsettled future for food and feed production and prices in general. Increasing demand, prices and fluctuations in supply in world markets for fishmeal, fish oil, soybean meal, maize and wheat meal emphasize the need to reduce the dependence of the fish feed industry on these ingredients by increasing choices among a wider range of raw materials. Legume seed such as peas, chickpeas and faba beans are promising ingredients for aquafeeds due to their high protein compared to cereals but also for their energy content. The objective of the present study was to investigate the use of these legumes in both high and low inclusion level in diets for the two main species farmed in the Mediterranean countries namely, European seabass (Dicentrarchus labrax L.) and gilthead seabream (Sparus aurata L.). In Chapter 3 the effects of different processing conditions were examined on whole seed flours of tested ingredients with respect to both nutritional and antinutritional factors and physical characteristics of the experimental diet pellets including high and low levels of each legume. Extrusion of raw material resulted in a clear reduction in trypsin inhibitors (TI) with chickpeas showing a decrease of up to 90% and complete inactivation for wheat flour, while for peas and faba beans reduction was less than 50% in most cases. Extrusion processing was less effective in the reduction of phytic acid and total tannins, occasionally reaching 22% and 18% respectively. Oligosaccharides and total NSP were not drastically affected by processing, however faba bean NSP showed greater reduction. A redistribution of soluble and insoluble NSP fractions was noted for chickpea and pea flours after extrusion. Physical characteristics of the pellets were not negatively affected for any of the tested diets. In Experiment I the effects of total or partial wheat substitution by legumes were investigated on nutrient digestibility, gastrointestinal evacuation rate and serum glucose response of European seabass. Use of legumes in seabass and seabream diets resulted in an overall increase in gastrointestinal evacuation time and a delay in glucose load. Specifically, gastric evacuation time was greatly delayed when seabass was fed a diet with high levels (30%) of chickpeas, while foregut evacuation time was mostly delayed by the diet including a high level (30%) of faba beans. In addition, glucose levels in seabass serum were also affected by the type of carbohydrates ingested with wheat starch showing more rapid increase and decrease of glucose compared to fish fed pea and chickpea diets, while faba bean starch resulted in a lower glucose peak. In Experiment II growth, digestibility, hematological parameters, histological effects and fillet organoleptic characteristics and the interaction between starch inclusion level (8% and 16% or 17% and 35% of legumes respectively) and legume type were estimated when tested legumes replaced wheat in European seabass diets. Digestibility coefficients were satisfactory for all nutrients (>93%) while legume diets at a low level had beneficial effects on growth parameters when compared to the control diet, with chickpeas showing a significant improvement in SGR (P<0.05). High level legume diets did not result in any negative effect on growth. HSI was increased with increasing starch/legume inclusion in the feed and serum glucose also increased for fish fed high levels of faba beans and chickpeas. Carcass proximate composition was not affected by replacement of wheat in the diets, excluding the increase of fat content in fish fed chickpeas. Sensory analysis showed no differences between fish fed the control and high legume inclusion diets. Lastly in Experiment III growth, hematological parameters, histological effects and the interaction between starch inclusion level (low and high) and legume type were evaluated when tested legumes were included in gilthead seabream diets. Decreased, but not significantly so, growth was observed for all diets including legumes compared to the control. Poorer SGR were observed for pea and faba bean diets when these legumes were included at high levels. Liver glycogen increased with increasing starch level, but HSI did not differ significantly for any of the diet treatments. Histological examination of hindgut did not show pathological effects, such as enteritis, for in either species or for any of the diets. Increased absorptive vacuoles were found for control and pea diets (high level) only for seabass. The findings of this thesis showed that the two important species cultivated in Mediterranean countries responded differently to the same raw materials used at high levels in the diets. Overall legumes had a strong effect on gastrointestinal evacuation reducing the rate of feed or digesta passage. Peas, chickpeas and faba beans successfully replaced wheat in seabass diets resulting in improved growth coefficients. However, when the same legumes included in seabream diets growth performance was not improved compared to the wheat based diet.

Lack of affordable feeds is one of the major constraints facing small-scale fish farmers in Tanzania. This study evaluated the suitability of moringa leaf meal (MLM), cassava leaf meal (CLM) and cassava root meal (CRM) as novel ingredients in Nile tilapia, Oreochromis niloticus diets. Each of the ingredients was processed in an attempt to remove the most significant antinutritional factor. A series of five experiments was conducted in a recirculation system using juvenile O. niloticus. The fish were fed isonitrogenous (30g 100g-1), isolipidic (10g 100g-1) and isoenergetic (18 kJ g-1) diets containing graded levels of the processed ingredients to their apparent appetite but not exceeding 10% of their body weight for a period of 8 weeks. Processing led to the removal of 0.3% of saponin from MLM and 60% and 90% of hydrogen cyanide from CLM and CRM respectively. The contents of other inherent antinutritional factors such as phenols, tannins and phytic acid were little affected. Processed MLM, CLM and CRM had 31.1/29.0/1.5g 100g-1 of crude protein, 5.9/10.2/2.4g 100g-1 of crude fibre and 20.1/19.7/15.8k Jg-1 of gross energy. The content of sulphur amino acids was higher in CLM (0.47%) than in MLM (0.23%). Digestibe protein and digestible energy was higher in MLM (25.71g 100g-1/15.44kJ g-1) than in CLM (12.71g 100-1/9.16kJ g-1). CRM had a digestible energy content of 13.5kJ g-1. Inclusion of either of the leaf meals, even at the lowest level of 15g 100g-1 of total dietary protein, led to a significant reduction in feed intake, growth and feed utilisation. Liver and small intestine did not show any histopathological changes which could be linked to dietary treatment. Conversely, cassava root meal could replace up to 75% of wheat meal in the diet without significantly affecting performance. The performance of leaf meals was marginally improved by a combination of blending and feeding stimulants, whereby a blend containing 1 part MLM and 2 parts CLM could provide up to 20g 100g-1 of dietary protein without significantly reducing performance. Biological and economic performance of practical diets containing 30-50g 100g-1 of dietary protein from moringa and cassava blends (LMB) with feeding stimulants was significantly lower than a fishmeal-meal based diet (FM) but comparable to a soybean meal-based diet (SBM). The suitability of MLM and CLM as novel protein sources in O. niloticus diets will depend on 1) improving reduction/removal of inherent antinutritional factors in MLM and CLM as well as improving digestibility of CLM. On the other hand, the suitability of CRM as a carbohydrate energy source will depend on the availability of cost effective protein sources due to its low protein content.

Traditionally, fish oil (FO) has been used extensively in aquafeeds. The stagnation in global fish oil production coupled with an increased demand for its use in aquaculture feeds, especially salmonid feeds, has greatly inflated fish oil prices. Therefore, in order to sustain the rapid growth of the tilapia industry, the dependence on these commodities in feeds should be reduced through use of cheaper and more sustainable sources of dietary lipids, such as palm oil. This study therefore investigated several, previously poorly understood, effects of palm oil on reproductive performance of the commercial tilapia species, Oreochromis niloticus; which currently ranks as second most popular species in world aquaculture. In the present study broodstock were fed on experimental diets at full and half ration regimes throughout their entire life cycle from exogenous feeding. Studies were conducted in standardised and controlled hatchery conditions, thereby reducing the potential influence of environmental variations. First feeding O. niloticus fry were fed on four diets, cod liver oil (D 1), palm oil (D 2), mixed palm and cod liver oil (D 3) (9:1 ration) and a commercial trout diet as control (D 4) (Skretting, U.K.) on a reducing ration based on fish size. The present study investigated the effect of dietary lipid sources on (1) growth performance, (2) biochemical composition of eggs (total lipid and fatty acid composition), (3) morphological parameters of eggs (total and relative fecundity, egg size, egg weight and EW:BW), (4) larval quality (larval length and weight) and (5) oocyte recruitment and its associated sex steroid hormones. Experimental diets and feeding ration significantly influenced (p<0.05) the growth performance over a period of 120 days. Total lipid and fatty acid composition of eggs originating from broodstock fed on palm oil, mixed palm and cod liver oil (9:1) or a control diet were not significantly different (P>0.05) when fed at either full (3% BWday-1) or half ration (1.5% BWday-1). The present study, however, confirmed that fatty acid composition of fish eggs reflected the fatty acid composition of the diet, although specific fatty acids were selectively utilized or retained in the eggs. The mean inter-spawning interval (ISI) increased with increasing fish size and averaged 14, 19 and 24 days for fish fed on palm oil, mixed palm and cod liver oil or control diets, respectively. The shortest ISI observed was 7 days for fish fed a palm oil diet. Total fecundity ranged from 660 - 820 eggs/clutch. Mean total fecundity was 750, 820 and 660 eggs/clutch for fish fed a palm, mixed palm and cod liver oil or a control diet, respectively, but these differences were not significant (P>0.05). However, relative fecundity and egg weight to body weight rates as a percentage (EW: BW) were found significantly differ (p<0.05) between fish fed the control diet and experimental diets. Mean egg diameter (2.2 mm) was not significantly influenced (p>0.05) by experimental diets. The egg volume, egg dry and wet weight, fertilisation and hatching rate were also not significantly different between fish fed the experimental diets. Oocyte development was classified into distinct stages based upon oocyte size, biochemical properties and structure. The recrudescence to these stages was not significantly influenced by broodstock fed experimental diets either at full or half ration. Steroid hormones and histological analyses provided valuable data concerning the oocyte development and recruitment in this species. Levels of 17ß-oestradiol (E2) and testosterone (T) peaked within 6 days of spawning, suggesting that vitellogenesis began as early as day 2 or 3 post-spawning. By day 6, ovaries were dominated by large late-vitellogenic/maturing oocytes (stages 6 & 7) occupying about 70% of the ovary. Gonadosomatic index (GSI) reached maximal levels by day 6. It is suggested that pre-vitellogenic oocytes are recruited into vitellogenic growth immediately after spawning and complete vitellogenesis on day 6 post-spawning. Finally, the present study investigated the effect of food restriction at two rations (full and half) on broodstock reproductive performance. Oreochromis niloticus were rationed from first feeding and throughout their life-cycle. The dietary regime, full ration (3%) and half ration (1.5%), influenced fish size but despite this variation no significant differences (p>0.05) were detected in total lipid and fatty acid composition in the eggs, total fecundity, egg diameter, total egg volume and larval size. These results suggested that despite large differences in food availability throughout their life cycle, investment in reproduction had remained remarkably consistent. It appeared that during food restriction, O. niloticus sacrificed body weight and growth so as to maintain reproductive investment. In summary, this study provides valuable information using a novel experimental design on the effects of dietary lipid sources on reproductive performance of female O. niloticus. Substituting palm oil for fish oil as the dietary lipid source and reducing ration by half (1.5% BWday-1) had no significant effect on reproductive performance. Therefore it is suggested that under controlled conditions, lipids of non-marine origin, such as palm oil, can be successfully substituted for broodstock diets. Halving feed requirement should also increase profitability of seed production. KEYWORDS: Tilapia; O. niloticus; palm oil; diet; fecundity; spawning periodicity; oocyte recruitment; reproductive performance.

Fish meal (FM) and fish oil (FO) have traditionally been central in aquaculture feed formulation but the finite global supply situation limiting future use along with issues of contaminant levels in these feed ingredients have become critical issues. The objectives of the present study were to investigate alternative feed ingredients as substitutes for both FM and FO in feeds for Atlantic salmon (Salmo salar) to ensure optimal growth, feed efficiency and health of the fish as well as maintaining the nutritional quality of the fish product to the human consumer, especially the levels of n-3 highly unsaturated fatty acid (HUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), in the flesh. The results of the present study revealed that there were no negative effects on growth performance, feed utilisation and apparent digestibility in Atlantic salmon when FO was substituted with vegetable oil (VO) but these parameters were affected when FM was replaced with alternative protein sources from plants and animals at high levels, despite dietary supplementation with crystalline amino acids and lecithin. Reduction in feed intake was a factor affecting growth retardation when FM inclusion decreased. However, replacing FM with alternative plant and animal proteins along with partial replacement of FO had no major effect on nutritional quality, particularly n-3 HUFA content of salmon tissues. Replacing Northern FO with decontaminated FO or blends of southern hemisphere FO and VOs strategies to reduce POP contaminants and retain high nutritional values in flesh were very successful. Dietary treatments and genetic origin of fish both had effects on tissue compositions and gene expression. All fish groups (strain/family), consist of CAL, LEAN and FAT strains, fed a diet containing VO showed significant differential expression of lipid metabolism-related genes compared to fish fed a FO diet with LEAN strain appearing to adapt to VO inclusion better than FAT strain. This thesis has demonstrated dual replacement of FM and FO with alternative raw materials in salmon feeds without a major negative impact on nutritional quality.

Intensive and semi-intensive aquaculture systems are dependent on nutrient input either in the form of supplemental or complete feeds. Most complete diets still include high fish meal (FM) levels (≥10%). However, as the industry attempts to reduce its reliance on FM, feeds must now be formulated with much lower levels especially for omnivorous species such as tilapia. By 2015, mean FM inclusion in tilapia diets was projected to fall below 3% and be further reduced to 1% by 2020. In the global context of competition for crops, finding suitable plant-based replacers for FM and meeting the increasing demand for seafood, lower-cost and under-utilised plant feedstuffs are now receiving greater attention. The study was divided into three distinct components – field survey, growth experiments, and life cycle assessment. Field surveys were used to contextualise the growth experiments and assess commercialisation opportunities for multi-enzyme inclusion in tilapia feeds. Two sets of digestibility and growth experiments were designed to evaluate the feasibility of using high inclusions of plant-based ingredients sourced from locally available feedstuffs in Thailand to substitute FM at low inclusion levels (0 – 5%). The research evaluated the hypothesis regarding the potential of exogenous enzymes (protease, xylanase and phytase) to minimize anti-nutritional effects on nutrient digestibility of proteins, polysaccharides and phosphorus in tilapia. The research also assessed the secondary effects of enzyme supplementation on economic efficiency and life cycle environmental impacts. Tilapia is the second most cultured finfish globally and Thailand is the sixth largest producer. Based on the findings of the field survey, feeding practices of Thai tilapia farmers were confirmed to be diverse. Feed inputs included, but were not limited to, agro-industrial by-products (e.g. rice bran, corn bran etc.) and commercial diets. Commercial diets contained 15 – 30% crude protein and lower protein livestock diets (i.e. pig ration) were often used for supplemental feeding or “fattening”. The experimental low FM diets were therefore formulated as grow-out or “fattening” diets for semi-intensive green-water systems, a prominent feature (>60%) of Thai tilapia farming. In Phase 1, the digestibility experiment assessed the digestibility and growth in tilapia fed 0%, 3% and 5% FM diets with and without xylanase (0.385 g kg-1) and phytase (0.075 g kg-1). Performance decreased significantly with declining FM levels. No differences in feed intake, feed conversion ratio (FCR), specific growth rate (SGR) and weight gain were observed between the enzyme and control diets. Nevertheless, tilapia fed the enzyme supplemented 3% FM and control 5% FM performed similarly (P < 0.05). No enzyme-related effects were noticed for protein digestibility but phosphorus (P) digestibility improved by 9%, except at 0% FM level (P > 0.05). The enzymes had no apparent influence on nitrogen (N) retention contrary to previous studies, however, higher retention for P was observed. Villus length decreased with declining FM levels yet no improvements were seen in tilapia fed enzyme diets. In a simultaneous grow-out experiment, the six experimental diets were compared to an industry 10% FM standard. Conversion ratio was the lowest (1.66) in adult tilapia fed 10% FM diet however the enzyme supplemented 0% FM fed fish had a comparatively low FCR of 1.67. There were no significant enzyme-related effects on weight gain¸ SGR and protein efficiency. Proximal villi results were inconsistent. The cost of feed decreased with declining FM levels but increased with enzyme inclusion. Nevertheless, the economic returns per kg of whole fish produced were better using enzyme supplemented diets compared to the controls. Though the size of the effects on growth and nutrient utilisation were modest, the findings suggested that xylanase and phytase had some level of synergistic action on the targeted anti-nutrients. However, further research was required. In Phase 2, two control diets (2% FM, negative control (NC) and 10% FM, positive control (PC)) were compared with three enzyme supplemented 2% FM diets (NO-PRO, 0.385 g kg-1 xylanase and 0.075 g kg-1 phytase only; LO-PRO, xylanase + phytase + 0.2 g kg-1 protease and HI-PRO, xylanase + phytase + 0.4 g kg-1 PRO). Growth performances improved with enzyme supplementation compared to the NC (P < 0.05). Of the enzyme supplemented diets, the LO-PRO diet showed the highest improvements in weight gain (26%) and feed intake (19%), the latter comparing statistically to the 10% FM PC diet. The HI-PRO diet had the best FCR (1.88), again comparable to the PC (1.73). The NO-PRO diet had the highest protein, P, lipid and energy digestibility, suggesting no additive effect of protease on these coefficients. In terms of gut histomorphology, the LO-PRO and PC diets had the highest measurements and were statistically similar which may have explained similarities in feed intake. Compared to the NC, the HI-PRO diet produced the highest level of change in net profit due to gains in feeding efficiency however, the LO-PRO showed better improvements in terms of growth. Based on these findings, the ternary combination of protease with xylanase and phytase (LOPRO) has potential in limiting FM use for tilapia grow-out feeds, however, the economic efficiencies were still below that of a 10% FM diet. Future considerations for research should target the indigestible dietary components in order to optimise enzyme dosages and maximise the benefits of each enzymes. In conclusion, a comparative life cycle assessment (LCA) was used to evaluate the environmental impacts of low FM diets and commercial feeds associated with tilapia production in Thailand. The study showed that the low FM enzyme supplemented diets had lower impact potentials and were environmental superior to the average (10% FM) commercial standard. LCA modules are recommended for least-cost formulation programmes as an option going forward. Additionally, LCA can be used as a predictive tool to guide farmers, especially small-scale producers, on the potential impacts of feed input choices and feeding practices. This will ensure higher product quality but also demonstrate environmental responsibility on the part of aquafeed and fish producers to final seafood consumers.

Current feed formulations within the aquaculture industry have tended to rely on high dietary lipid thus offsetting relatively expensive protein as a source of energy. In this way, protein can be ‘spared’ for synthesis of new tissue and the high lipid content can also fulfil both fish and consumer essential fatty acid (EFA) requirements. However, the main disadvantage of feeding high lipid levels to farmed fish is a surplus of fat deposition in the flesh and other important tissues, which can detrimentally impact on quality characteristics central to the human consumer. However, based on previous work in other animal models, it is entirely feasible that supplementation of the diet with bioactive fatty acids such as conjugated linoleic acid (CLA) and tetradecylthioacetic acid (TTA) may mitigate the deleterious effects of feeding farmed fish high fat diets by reducing fat deposition in particular. The general objective of this research work was to test the hypothesis that CLA and/or TTA could augment growth, reduce fat deposition and enhance fatty acid composition via incorporation of these bioactive fatty acids, and increase n-3 highly unsaturated fatty acid (HUFA) levels in the flesh of commercially important fish species such as Atlantic salmon (Salmo salar), Atlantic cod (Gadus morhua L.) and rainbow trout (Oncorhynchus mykiss). This project also considered the influence of CLA and TTA on enzymes and transcription factors thought to be pivotal in lipid metabolism and fatty acid oxidation in particular. A subsidiary aim of this research work was to investigate the immunological impact of dietary CLA and TTA administration in these fish. The results of this project have revealed that the hypothesis was only partly proved. There was no effect in growth or biometry after either CLA or TTA supplementation in any of the fish species investigated. Additionally, there were few physiologically significant effects on fat levels on fish as a result of TTA or CLA administration. However, there were a number of effects on fatty acid metabolism including inhibition of steroyl coenzyme desaturase (SCD) in cod and trout in particular and also enhancement of hepatic n-3 HUFA levels in trout. Importantly, it was determined that both TTA and CLA could be incorporated into the flesh thus providing a vehicle through which these bioactive fatty acids can be delivered to the consumer. There were also a number of beneficial effects on activity and gene expression of a number of enzymes and transcription factors thought to be fundamental to the modulation of fatty acid oxidation in particular. However, the effects on gene transcription and biochemistry had little impact at the whole body level. This research work also showed that there were no detrimental effects on immune status after supplementation with dietary CLA or TTA. Conclusively, this thesis has contributed to the overall understanding of the influence of dietary CLA and TTA in farmed fish.

Ma Wing-chi, Joyce. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 131-148). / Abstracts in English and Chinese. / Abstract (in English) --- p.i / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.vi / Contents --- p.viii / Abbreviation --- p.xii / List of Tables --- p.xiii / List of Figures --- p.xv / Chapter 1. --- Introduction --- p.1 / Chapter 2. --- Literature review --- p.5 / Chapter 2.1 --- Fisheries in Hong Kong --- p.5 / Chapter 2.2 --- Flavor of seafood --- p.6 / Chapter 2.2.1 --- Lipid-derived volatile aroma compounds --- p.7 / Chapter 2.2.2 --- "Alcohols, aldehydes and ketones" --- p.8 / Chapter 2.2.3 --- Enzymatic conversion of sulfur- and nitrogen-containing precursors --- p.9 / Chapter 2.2.4 --- Thermally generated compounds --- p.9 / Chapter 2.2.5 --- Bromophenols --- p.10 / Chapter 2.2.5.1 --- General properties of bromophenols --- p.11 / Chapter 2.2.5.2 --- Threshold of bromophenols --- p.14 / Chapter 2.2.5.3 --- Toxicity of bromophenols --- p.17 / Chapter 2.2.5.4 --- Previous studies about bromophenols --- p.19 / Chapter 2.2.5.5 --- Bromophenols in aquacultured seafood --- p.20 / Chapter 2.2.5.6 --- Possible dietary sources of bromophenols --- p.20 / Chapter 2.2.5.7 --- Possibility of increasing bromophenol content in aquacultured fish --- p.23 / Chapter 2.3 --- Criteria for selecting experimental fish model --- p.24 / Chapter 3. --- Distribution of Bromophenols in selected Hong Kong seafoods --- p.27 / Chapter 3.1 --- Introduction --- p.27 / Chapter 3.2 --- Materials and methods --- p.28 / Chapter 3.2.1 --- Sample collection and preparation --- p.28 / Chapter 3.2.2 --- Simultaneous steam distillation-solvent extraction (SDE) --- p.30 / Chapter 3.2.3 --- Gas chromatography / mass spectrometry (GC/MS) --- p.30 / Chapter 3.2.4 --- Compound identification and quantification --- p.31 / Chapter 3.2.5 --- Recoveries --- p.33 / Chapter 3.2.6 --- Moisture determination --- p.34 / Chapter 3.2.7 --- Statistical analysis --- p.34 / Chapter 3.3 --- Results and discussion --- p.34 / Chapter 3.3.1 --- Distribution of bromophenols in seafoods --- p.34 / Chapter 3.3.1.1 --- Bromophenols in marine fishes --- p.49 / Chapter 3.3.1.2 --- Bromophenols in mollusks --- p.49 / Chapter 3.3.1.3 --- Bromophenols in crustaceans --- p.50 / Chapter 3.3.2 --- Seasonal variations of TBCs --- p.51 / Chapter 3.3.3 --- Bromophenols in diet contents --- p.52 / Chapter 3.3.4 --- Bromophenol contents of freshwater fish --- p.53 / Chapter 3.3.5 --- Relationship between the living habitats and bromophenol contents --- p.56 / Chapter 3.3.6 --- Bromophenols as flavor compounds in seafoods --- p.58 / Chapter 3.4 --- Conclusion --- p.59 / Chapter 4. --- Distribution of Bromophenols in selected Hong Kong seaweeds --- p.61 / Chapter 4.1 --- Introduction --- p.61 / Chapter 4.2 --- Materials and methods --- p.62 / Chapter 4.2.1 --- Sample collection and preparation --- p.62 / Chapter 4.2.2 --- Simultaneous steam distillation-solvent extraction (SDE) --- p.63 / Chapter 4.2.3 --- Gas chromatography / mass spectrometry (GC/MS) --- p.64 / Chapter 4.2.4 --- Compound identification and quantification --- p.65 / Chapter 4.2.5 --- Recoveries --- p.66 / Chapter 4.2.6 --- Moisture determination --- p.67 / Chapter 4.3 --- Results and discussion --- p.67 / Chapter 4.3.1 --- Distribution of bromophenols in marine algae --- p.67 / Chapter 4.3.2 --- Seasonal variations --- p.76 / Chapter 4.3.3 --- Functions of bromophenols in marine algae --- p.79 / Chapter 4.3.4 --- Marine algae as sources of bromophenols in marine environment --- p.80 / Chapter 4.4 --- Conclusion --- p.81 / Chapter 5. --- Enhancement of bromophenol contents in aquacultured fish by the development of bromophenol-rich fish feeds --- p.83 / Chapter 5.1 --- Introduction --- p.83 / Chapter 5.2 --- Materials and methods --- p.85 / Chapter 5.2.1 --- Preparation of fish feeds --- p.85 / Chapter 5.2.2 --- Storage conditions of fish feeds --- p.88 / Chapter 5.2.3 --- Experimental animals --- p.88 / Chapter 5.2.4 --- Solvent and chemicals --- p.90 / Chapter 5.2.5 --- Extraction and quantification of bromophenols --- p.90 / Chapter 5.2.5.1 --- Simultaneous steam distillation-solvent extraction (SDE) --- p.90 / Chapter 5.2.5.2 --- Gas chromatography / mass spectrometry (GC/MS) --- p.91 / Chapter 5.2.5.3 --- Compound identification and quantification --- p.92 / Chapter 5.2.5.4 --- Recoveries --- p.93 / Chapter 5.2.6 --- Moisture determination --- p.94 / Chapter 5.2.7 --- Statistical analysis --- p.94 / Chapter 5.2.8 --- Sensory test --- p.95 / Chapter 5.3 --- Results and discussion --- p.96 / Chapter 5.3.1 --- Bromophenol contents in wild-harvested and aquacultured fish --- p.96 / Chapter 5.3.2 --- Development of bromophenol-rich fish feed --- p.99 / Chapter 5.3.3 --- Effect of feeding the fish with the fish feed developed --- p.105 / Chapter 5.3.4 --- Sensory evaluation on the flesh of the fish fed with different fish feeds --- p.121 / Chapter 5.3.5 --- Growth of the fish fed with different fish feeds --- p.124 / Chapter 5.4 --- Conclusion --- p.126 / Chapter 6. --- General conclusion and significance of the study --- p.128 / References --- p.131