Effect of docosahexaenoic acid enrichment in Artemia on growth of Pacific bluefin tuna Thunnus orientalis larvae

Feeding enriched Artemia induces growth failure in Pacific bluefin tuna (PBT) Thunnus orientalis larvae; however, feeding of yolk-sac larvae of marine fish promotes larval growth. It is considered that this growth failure partly results from dietary docosahexaenoic acid (22:6n-3, DHA) deficiency. Therefore, we examined the effect of DHA contents in enriched Artemia on the growth of PBT larvae. Artemia nauplii were enriched with graded levels of DHA ethyl ester, and fed to PBT larvae for 9 days. Yolk-sac larvae of Japanese parrotfish Oplegnathus fasciatus were used as a reference diet. The DHA contents in Artemia increased with the enrichment from 0 mg g^− 1 dry weight basis (DW) to 25 mg g^− 1 DW, while the content in the reference diet was 21 mg g^− 1 DW. Feeding of enriched Artemia significantly improved the growth of PBT larvae. However, this improvement was negligible when compared with the excellent growth of the larvae that were fed the reference diet. PBT larvae that were fed the reference diet accumulated approximately twice or much higher levels of DHA in the neutral and polar lipids in the body when compared with the larvae that were fed enriched Artemia. These results show that PBT larval growth cannot be promoted by feeding enriched Artemia even if the DHA contents in Artemia are elevated to the same levels as those of yolk-sac larvae. The incorporation of dietary DHA into phospholipids in the fish body may be desirable for the normal growth of PBT larvae.     

Energetic physiology of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus fed with different microalgal diets

The Caribbean scallops Argopecten nucleus and Nodipecten nodosus are currently being placed into mass culture in Colombia. The limited availability of wild seed upon which to base these cultures has promoted research into the development of artificial production of this seed in hatcheries. In support of this effort, we studied the effects of different diets on the physiology of the two scallop species in order to determine the optimal feeding regimes for maintenance of adult specimens in the laboratory. Seven monoalgal diets were tested, including Chaetoceros calcitrans, Chaetoceros muelleri, Isochrysis galbana (Ig), Nannochloris oculata (No), Phaeodactylum tricornutum (Pt), Tetraselmis chui (Tc) and Tetraselmis tetrahele (Tt). Four mixed diets were also tested, including I. galbana + C. calcitrans (Ig + Cc), I. galbana + N. oculata (Ig + No), I. galbana + T. tetrahele (Ig + Tt), and I. galbana + lipid emulsion of docosahexanoic acid DHA (Ig + lip). All the dietary trials were carried out under uniform conditions of temperature (25 °C), salinity (36‰) and algal concentration (0.45 mg L^− 1). Physiological variables measured in association with each diet included feeding rates (clearance, ingestion and absorption), oxygen consumption and ammonium excretion rates as well as their scope for growth. The results showed that the best scope for growth for both scallops was obtained with diet Ig since this diet induced the highest feeding rates, accompanied by the lowest oxygen consumption and ammonium excretion. The feeding rates and scope for growth of A. nucleus were greater than those of N. nodosus for the majority of the diets, which was attributed to a higher rate of water pumping by the former species. Greater capacity for branchial food retention by A. nucleus was discarded as a possibility since N. nodosus had a greater branchial surface area per unit dry weight than A. nucleus. In spite of these differences, the oxygen consumption and the excretion rates were similar between the two scallops which suggested that A. nucleus was more efficient in its use of oxygen and retention of body proteins for physiological functioning. Mixed diets or addition of DHA did not permit increases in scope for growth in either of the scallops over that observed using monoalgal diet Ig, which suggest that biologically and economically this diet is optimal for the feeding of adult scallops in the laboratory.     

Comparison of early life history stages of the bay scallop, Argopecten irradians: Effects of microalgal diets on growth and biochemical composition

The culture of bay scallops, Argopecten irradians, is limited by a reliable and affordable supply of spat and the ability to ensure that animals attain market size within a single growing season. The main goals of our study were thus: (1) to develop growth-optimizing algal diets for implementation in hatcheries, and (2) to identify and compare bay scallop postlarval and juvenile dietary requirements, especially of lipids and fatty acids, which if met may enhance production. Nutritional needs of postlarval bay scallops (present study) are compared with those of sea scallops, Placopecten magellanicus, offered the same diets in a previous companion study. To this end, postlarval (initial shell height, SH = 240 μm) and juvenile (initial SH = 10 mm) bay scallops were offered 6–7 microalgal diet combinations at 20 °C, for 3 weeks. A similar growth ranking among diets was observed between the two developmental stages. A combination diet of Pavlova sp. (CCMP 459) and Chaetoceros muelleri was far superior to any other diet tested, yielding growth rates of 58 and 357 μm day^− 1 which were 65% and 25% higher than the next highest performing diet of Tetraselmis striata/C. muelleri in postlarvae and juveniles, respectively. The T. striata/C. muelleri diet, which is limited in the n-3 fatty acid docosahexaenoic acid (DHA), yielded very poor growth of sea scallop postlarvae in a prior study, indicating that bay scallops may have less stringent requirements for DHA than sea scallops. The Pav 459/C. muelleri diet, which also supported the highest growth of sea scallop postlarvae, is characterized by elevated levels of the n-6 fatty acids, arachidonic (AA) in C. muelleri and 4,7,10,13,16-docosapentaenoic (DPA) in Pav 459. The two diets deficient in AA and n-6 DPA, Pavlova lutheri/Thalassiosira weissflogii and P. lutheri/Fragilaria familica, yielded the lowest growth rates in both bay scallop postlarvae and juveniles. Tissue enrichment of these two fatty acids relative to the diet, as well as overall enrichment in ∑n-6 fatty acids was observed across developmental stages and dietary treatments. A similar pattern has previously been observed in sea scallop postlarvae, suggesting a dietary requirement for n-6 fatty acids in pectinids that has often been overlooked in the past.     

Pigmentation, carotenoids, lipid peroxides and lipid composition of skin of red porgy (Pagrus pagrus) fed diets supplemented with different astaxanthin sources

A feeding experiment was carried out to determine the efficiency of different commercial sources, chemical forms and levels, of dietary astaxanthin, to appropriately pigment the red porgy (Pagrus pagrus) skin. According to this, total carotenoid content, profiles and chemical forms present in the skin were determined. In order to establish the potential for antioxidant protecting role of astaxanthin supplemented diets, peroxide levels and lipid composition of skin were also determined.

Red porgy alevins were fed six dietary treatments in triplicate; a basal diet (B) without carotenoids; two diets (N25 and N50) formulated to supply either 25 or 50 mg kg^− 1 of an esterified source of astaxanthin (Haematococcus pluvialis, NatuRose™); two diets (CP25 and CP50) with either 25 or 50 mg kg^− 1 of unesterified astaxanthin (Carophyll® Pink); and a positive control diet (B + S) proved as a successful pigmenting-diet in previous experiences (B + S, 88% basal diet:12% frozen shrimp) [Cejas, J., Almansa, E., Tejera, N., Jerez, S., Bolaños, A., Lorenzo, A., 2003. Effect of dietary supplementation with shrimp on skin pigmentation and lipid composition of red porgy (P. pagrus) alevins. Aquaculture 218, 457–469].

All fish fed carotenoid supplemented diets displayed a pink-coloured skin after 4 months of feeding in contrast to the greyish appearance displayed by fish fed the basal diet not supplemented with carotenoids (B). Furthermore, astaxanthin diesters were the major carotenoid in the skin of pink fish. A second carotenoid, tentatively identified as tunaxanthin diester, was also detected. The best results in terms of skin natural reddish hue, total carotenoid and astaxanthin contents were found by using the esterified forms of dietary astaxanthin (N25, N50 and B + S). Interestingly, the lowest levels of lipid peroxides were found in the fish fed these three treatments. However, no effect of treatment on lipid composition was found. In conclusion, red porgy alevins are able to efficiently utilise dietary natural or synthetic astaxanthin, and deposit this pigment in its esterified form to acquire an acceptable pink-coloured skin compared to that of the wild fish.     

Atlantic salmon, Salmo salar, utilizes wax ester-rich oil from Calanus finmarchicus effectively

Against a background of decreasing availability of fish oils for use in aquaculture, the present study was undertaken to examine whether a wax ester-rich oil derived from the calanoid copepod Calanus finmarchicus, could be used effectively by Atlantic salmon when supplied in their diet. Individually tagged Atlantic salmon of initial weight around 500 g were divided into replicate tanks of two dietary groups and fed either a fish oil supplemented diet, or an experimental diet coated with Calanus oil. Wax esters accounted for 37.5% of the lipids in the Calanus oil diet but were absent from the fish oil diet in which triacylglycerols (TAG) were the major lipid class. Over the feeding period (140 days) the salmon fed fish oil displayed a greater increase in length, but there was no significant difference between the two groups in weight gained. The specific growth rates (0.75) and the feed conversion ratio of fish fed the two diets were similar throughout the study. No differences were observed in the apparent digestibility coefficients (ADC) of fish fed Calanus oil or fish oil. The ADC of fatty acids decreased with chain length and increased with unsaturation. Long-chain alcohol utilization showed a similar tendency although there was a notable difference in that saturated long-chain alcohols were utilized better than the comparable fatty acid homologue. In fecal lipid of fish fed Calanus oil, the content of 16:0 alcohol decreased in both the free long-chain alcohol and wax ester fractions, while the corresponding fatty acid increased in the feces of both dietary groups of fish. In contrast, the proportion of the 22:1n−11 alcohol increased in both fecal wax esters and free long-chain alcohol fractions whereas 22:1n−11 fatty acid displayed no accumulation. The observed patterns of fatty acid and long-chain alcohol compositions in fecal lipid compared to those of the initial dietary lipid are consistent with the digestive lipases of salmon preferentially hydrolyzing esters containing polyunsaturated fatty acid (PUFA) moieties. The wax esters of Calanus oil contained substantial amounts of the n−3 PUFA, 20:5n−3 and 22:6n−3, that were effectively deposited in muscle and liver tissues. No major differences were seen in either lipid content/lipid classes or in gross fatty acid composition of these tissues between the two dietary groups. It is concluded that that Atlantic salmon in seawater can effectively utilize diets in which a major lipid component is derived from zooplankton rich in wax ester without any detrimental change in growth or body lipid composition. This finding gives support to the use of lipid from zooplankton from high latitudes as an alternative or as a supplement to fish oil and a provider of long-chain n−3 PUFA in diets for use in salmon aquaculture.     

Immunization of grouper, Epinephelus coioides, confers protection against a protozoan parasite, Cryptocaryon irritans

The present study aimed to determine whether protection is conferred by immunization of grouper, Epinephelus coioides, against a protozoan parasite, Cryptocaryon irritans. The immunization of E. coioides was carried out by a low level exposure of fish to live C. irritans theronts from predetermined number of tomonts and by an intraperitoneal injection of a vaccine consisting of formalin-killed C. irritans theronts.

Mucus titers detected by ELISA were significantly higher in fingerling and adult grouper subjected to the low level of exposure to C. irritans theronts at 3-week post-exposure compared to fish that had no previous exposure. In addition, significantly smaller tomonts were produced from adult grouper after three successive exposures than the tomonts produced after a single exposure to the parasite.

In the vaccine-immunization experiment, no mortality was monitored in fish that received high dose vaccine (100 μg/fish), while 40% cumulative mortality and 100% cumulative mortality were recorded in low dose group (10 μg/fish) and control group (PBS-injected), respectively. In the succeeding replicate, the vaccine-immunized group (high dose) had 37.5% cumulative mortality and 100% cumulative mortality for the control. In addition, a total of 1830 tomonts were collected at 5-day post-challenge from the control group while none from the vaccine-immunized group. Significantly fewer trophonts and tomonts were enumerated at 5-day and 7-day post-challenge, respectively, in the vaccine-immunized group than the control.

Results suggest that a protective immunity has been conferred on the immunized grouper as indicated by high antibody titers in the mucus of C. irritans-exposed fish and higher survival and fewer parasites in vaccine-immunized fish than the control groups. The conferred immunity played a major role in preventing or limiting the adhesion, invasion, and development of C. irritans theronts on the skin of the immunized grouper.     

Effect of variation in the protein value of the red macroalga Palmaria palmata on the feeding, growth and gonad composition of the sea urchins Psammechinus miliaris and Paracentrotus lividus (Echinodermata)

The sea urchins Psammechinus miliaris and Paracentrotus lividus were fed three macroalgal diets with varying protein content over a 90-day period. These diets were the red alga Palmaria palmata, which had been grown in seawater enriched with two levels of ammonia-N and contained a protein content of either 41% (High-N) or 32% (Low-N) and the brown alga Laminaria saccharina (23% protein). A significantly different response to the macroalgal diets by the two sea urchin species was found in terms of food consumption rate, food conversion efficiency, gonadal growth and gonad nutritional composition. P. miliaris was able to efficiently utilise the High-N P. palmata, whereas P. lividus exhibited a higher food conversion efficiency (FCE) and specific growth rate (SGR) when fed the Low-N compared with the High-N P. palmata. P. miliaris had a significantly higher food consumption rate and higher gonad index (GI) compared with P. lividus, irrespective of diet type. The lowest FCE, SGR and GI were shown by both species fed L. saccharina. No interspecific difference in gonadal nitrogen content was observed, although a positive relationship was shown between dietary and gonadal nitrogen content for both species. All three macroalgal diets produced bright orange or yellow orange gonad colour in P. lividus. High-N and Low-N P. palmata improved gonad colouration in P. miliaris compared with L. saccharina. The results suggest that P. miliaris can efficiently assimilate high protein plant-based diet, whereas P. lividus is less capable of benefiting from high dietary protein levels and this should be taken into consideration when culturing this species either in land-based or sea-based polyculture systems.     

Artemia enriched with high n-3 HUFA may give a large improvement in performance of Atlantic halibut (Hippoglossus hippoglossus L.) larvae

Atlantic halibut larvae were fed Artemia enriched with two different oil emulsions (cod liver oil and 2050TG) from first feeding to 70 days after first-feeding (dpff). Larvae fed 2050TG enriched Artemia had better growth, survival and eye migration than larvae fed the cod liver oil enriched Artemia, while pigmentation rate was similar in the two groups. In addition to the difference in fatty acids, the two emulsions differed in lipid class composition, since 2050TG is a synthetic oil and a mixture of mono-, di- and tri-acylglycerol, while cod liver oil is a tri-acylglycerol. Total lipid level, estimated as fatty acid methyl esters (FAME) was similar in the two Artemia types, but sum of n-6 and n-3 fatty acids, arachidonic acid (20:4n-6, ARA), docosahexaenoic acid (22:6n-3, DHA) and eicosapentaenoic acid (20:5n-3, EPA) were higher in Artemia enriched with 2050TG than in the cod liver oil enriched Artemia. However, the main difference in fatty acid composition in the larvae, was a higher DHA (% of total fatty acids) in 2050TG larvae than in cod liver oil larvae. The lipid level measured as FAME was up to four times higher in the 2050TG larvae than in the cod liver oil larvae, and the reason for this may have been a better bioavailability of the partly digested lipid in the 2050TG emulsion. The correlation between a high level of lipid in the larval tissues (e.g. high energy status) and improved eye migration in larvae fed the 2050TG enriched Artemia supports the hypothesis that energy limitation on the larval stage may be a cause of the impaired eye migration commonly observed in farmed Atlantic halibut juveniles.     

Comparison of effects of vegetable oils blended with southern hemisphere fish oil and decontaminated northern hemisphere fish oil on growth performance, composition and gene expression in Atlantic salmon (Salmo salar L.)

Replacement of fish oil with sustainable alternatives, such as vegetable oil, in aquaculture diets has to be achieved without compromising the nutritional quality, in terms of n-3 highly unsaturated fatty acid (HUFA) content, of the product. This may be possible if the level of replacement is not too high and oil blends are chosen carefully but, if high levels of fish oil are substituted, a fish oil finishing diet prior to harvest would be required to restore n-3HUFA. However, a decontaminated fish oil would be required to avoid increasing undesirable contaminants. Here we test the hypotheses that blending of rapeseed and soybean oils with southern hemisphere fish oil will have a low impact upon tissue n-3HUFA levels, and that decontamination of fish oil will have no major effect on the nutritional quality of fish oil as a feed ingredient for Atlantic salmon. Salmon (initial weight ~ 0.8 kg) were fed for 10 weeks with diets in which 60% of fish oil was replaced with blends of soybean, rapeseed and southern hemisphere fish oil (SVO) or 100% decontaminated northern fish oil (DFO) in comparison with a standard northern fish oil diet (FO). Decontamination of the oil was a two-step procedure that included treatment with activated carbon followed by thin film deodorisation. Growth performance and feed efficiency were unaffected by either the SVO or DFO diets despite these having lower gross nutrient and fatty acid digestibilities than the FO diet. There were also no effects on the gross composition of the fish. Liver and, to a lesser extent flesh, lipid levels were lower in fish fed the SVO blends, due to lower proportions of neutral lipids, specifically triacylglycerol. Tissue lipid levels were not affected in fish fed the DFO diet. Reflecting the diet, flesh eicosapentaenoic acid (EPA) and total n-3 fatty acids were higher, and 18:1n-9 lower, in fish fed DFO than FO, whereas there were no differences in liver fatty acid compositions. Flesh EPA levels were only slightly reduced from about 6% to 5% although docosahexaenoic acid (DHA) was reduced more severely from around 13% to about 7% in fish fed the SVO diets. In contrast, the liver fatty acid compositions showed higher levels of n-3 HUFA, with DHA only reduced from 21% to about 18% and EPA increased from under 8% to 9–10% in fish fed the SVO diets. The evidence suggested that increased liver EPA (and arachidonic acid) was not simply retention, but also conversion of dietary 18:3n-3 and 18:2n-6. Increased HUFA synthesis was supported by increased hepatic expression of fatty acyl desaturases in fish fed the SVO diets. Flesh n-3HUFA levels and desaturase expression was significantly higher in fish fed soybean oil than in fish fed rapeseed oil. In conclusion, partial replacement of fish oil with blends of vegetable oils and southern hemisphere fish oil had minimal impact on HUFA levels in liver, but a greater effect on flesh HUFA levels. Despite lower apparent digestibility, decontamination of fish oil did not significantly impact its nutritional quality for salmon.     

Effects of six alternative lipid sources on growth and tissue fatty acid composition in Japanese sea bass (Lateolabrax japonicus)

Ten-week experiment was carried out on Japanese sea bass (5.87 ± 0.02 g) to study the effects of replacement of fish oil with six alternative lipid sources: pork lard, PL; beef tallow, BT; poultry fat, PF; soybean oil, SO; corn oil, CO; and a mixed-fat (MF: tallow, 60%; soy oil, 20%; fish oil, 20%) on growth performance and fatty acid (FA) composition in fillet and liver. Seven isoenergetic and isonitrogenous experimental diets were formulated, containing 10% of added lipid. Fish oil was used in control diet, which was substituted by 50% with the alternative lipid sources in the other six diets.

Weight gain (WG), specific growth rate (SGR), Feed conversion ratio (FCR) feed intake and hepatosomatic index (HSI) of fish fed the experimental diets were not significantly different (P > 0.05). Protein efficiency ratio (PER) in fish fed the PF diet were significantly lower than those of fish fed SO and CO diets. Significant differences in carcass moisture and lipid contents of carcass and liver were observed among fish fed the dietary treatments. Generally, the fatty acid composition of fish fillets and livers reflected the dietary FA composition.     

The utilization of a filamentous green alga (Cladophora glomerata (L) Kutzin) as a protein source in pelleted feeds for Sarotherodon (Tilapia) niloticus fingerlings

Duplicate groups of S. niloticus fingerlings were fed each of five different diets for 8 weeks. The diets contained 30% crude protein supplied by varying proportions of fishmeal and C. glomerata meal. The five diets were formulated to supply fishmeal protein: C. glomerata meal protein ratios of 30 : 0 (diet 1), 25 : 5 (diet 2), 20 : 10 (diet 3), 15 : 5 (diet 4) and 10 : 20 (diet 5) respectively. A sixth diet (diet 6) containing 25% crude protein supplied entirely by C. glomerata meal was also fed.

Diets 1 and 2 produced significantly the highest growth rates and best protein utilization. Highest values for protein digestibility were found in diets 2 and 3 (94.5 and 93.9% respectively) although the values were not significantly different from other treatments. Weight gain and protein utilization decreased as the level of algal protein increased in diets 2 to 6. Carcass analysis revealed a decrease in lipid and protein contents with increasing levels of algae in the diet. It is postulated that algal meals are viable partial dietary protein sources for tilapia.     

Alternative vegetable lipid sources in diets for grouper, Epinephelus coioides (Hamilton): effects on growth, and muscle and liver fatty acid composition

The aim of this study was to investigate the effects of different oils on growth performance and lipid metabolism of the grouper, Epinephelus coioides. Five experimental fish meal‐based isonitrogenous and isolipidic diets were formulated containing either 5.5%‐added fish oil (FO), soybean oil (SBO), corn oil (CO), sunflower oil (SFO) or peanut oil (PO). Each diet was fed to triplicate groups of 20 fish (initial body weight 13.2±0.02 g) grown in seawater at 28.0–30.5 °C for 8 weeks. Fish were fed twice a day to visual satiety. No significant differences in the survival, weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio or hepatosomatic index were found between fish fed the FO or vegetable oils (VO) diets. Dietary lipid sources did not affect whole‐body composition among grouper fed the various diets. Muscle of fish fed the FO diet had significantly higher levels of 14:0, 16:0, 16:1n‐7, 20:5n‐3[eicosapentaenoic acid (EPA)] and docosahexaenoic acid (DHA)+EPA (except for PO fed fish) compared with those of fish fed VO diets. However, the levels of 18:1n‐9, 18:2n‐6 and DHA/EPA ratios in the muscle of fish fed FO diet were significantly lower than those of fish fed the VO diets. The liver of fish fed the FO diet had significantly higher levels of 18:0, 20:5n‐3, 22:6n‐3, n‐3 highly unsaturated fatty acids and DHA+EPA than those of fish fed the VO diets, whereas increases in 18:1n‐9, 18:2n‐6 and mono‐unsaturated fatty acid levels were observed in the liver of fish fed the VO diets.     

Effect of fish-meal replacement with poultry by-product meal on the growth, tissue composition and hematological parameters of largemouth bass (Micropterus salmoides) fed diets containing different lipids

We conducted a 12-week feeding trial with largemouth bass (LMB) to examine the interactive effects of non-fish dietary proteins with different lipid sources on the performance of LMB (3.4 g ± 0.05 g). Practical diets contained poultry by-product meal (PBM) in place of fish meal on a digestible-protein basis, and 12% blood meal (BM). Diets were supplemented with 10% lipid as canola (CAN), chicken (CHK), CHK + MFO (50 : 50), menhaden fish oil (MFO) or CHK + CAN (50:50). A commercial trout diet (SC) was included as a positive control. Growth, survival, feed intake (FI), feed conversion ratio (FCR), protein efficiency ratio (PER), body composition, fatty acid composition of liver and muscle, hematological parameters and lysozyme and alternative complement activity (ACH50) were measured to assess diet effects. Weight gain (23.8 ± 1.8 g), survival (92 ± 4%), FI (44.3 ± 4.1 g), FCR (2.1 ± 0.1) and PER (1.3 ± 0.04) of LMB did not differ among the replacement diets. However, weight gain (38.5 ± 1.5), FI (53.9 ± 1.1), and PER (1.6 ± 0.1) of LMB fed the SC diet was higher and FCR (1.4 ± 0.1) was lower than that of fish fed the test diets. No signs of essential fatty acid deficiency were observed, although tissue concentrations of n-3 highly unsaturated fatty acids declined markedly in LMB fed diets without fish oil. Lymphocytes were elevated in fish fed diets with ≥ 7% n-3 fatty acids. Fish fed diets with 10% MFO or CHK + MFO had higher ACH50 than fish fed the diet with 10% CHK. There were no differences in serum lysozyme activity among fish fed the test diets. Hematocrit, hemoglobin, ACH50, and lysozyme activity were higher in fish fed the SC diet than in those fed the test diets. Regardless of lipid source, the ACH50 and lysozyme activity were greatly reduced in LMB fed PBM diets compared to LMB fed diets with 30% fish meal in a previous trial. The amino acid composition of the PBM was similar to that of fish meal. However, the availability of some essential amino acids from PBM to LMB may have been limited, and poor diet palatability caused by BM also may have reduced feed intake and performance of LMB.     

Replacement of fish meal by meat and bone meal in diets for large yellow croaker, Pseudosciaena crocea

A growth experiment was conducted to investigate the effects of replacement of fish meal (FM) by meat and bone meal (MBM) in diets on the growth and body composition of large yellow croaker (Pseudosciaena crocea). Six isonitrogenous (43% crude protein) and isoenergetic (20 kJ g^− 1) diets replacing 0, 15, 30, 45, 60 and 75% FM protein by MBM protein were formulated. Each diet was randomly allocated to triplicate groups of fish in sea floating cages (1.0 × 1.0 × 1.5 m), and each cage was stocked with 180 fish (initial average weight of 1.88 ± 0.02 g). Fish were fed twice daily (05:00 and 17:30) to apparent satiation for 8 weeks. The water temperature ranged from 26.5 to 32.5 °C, salinity from 32 to 36‰, and dissolved oxygen content was approximately 7 mg l^− 1 during the experimental period. Survival decreased with increasing dietary MBM and the survival in the fish fed the diet with 75% protein from MBM was significantly lower than other groups (P < 0.05). There were no significant differences in specific growth rate (SGR) among the fish fed the diets with 0 (the control group), 15, 30 and 45% protein from MBM. However, SGR in the fish fed the diets with 60 and 75% protein from MBM were significantly lower than other groups (P < 0.05). No significant differences in feeding rate were observed among dietary treatments. The digestibility experiment showed that the apparent digestibility coefficients (ADC) of dry matter, protein, lipid and energy of MBM were significantly lower compared with those of FM (P < 0.05). Essential amino acid index was found to be correlated positively with SGR in the present study, suggesting that essential amino acid balance was important. Body composition analysis showed that the carcass protein and essential amino acids were not significantly affected by dietary MBM. The lipid and n-3 highly unsaturated fatty acid (n-3 HUFA) in fish muscle, however, significantly decreased with increasing dietary MBM. These results showed that 45% of FM protein could be replaced by MBM protein in diets of large yellow croaker without significantly reducing growth. It was suggested that the reduced growth with higher MBM was due to lower digestibility and imbalance of essential amino acids.     

Effects of dietary canola oil level on growth, fatty acid composition and osmoregulatory ability of juvenile fall chinook salmon (Oncorhynchus tshawytscha)

This study assessed refined canola oil (CO) as a supplemental dietary lipid source for juvenile fall chinook salmon, Oncorhynchus tshawytscha, parr with respect to possible effects on their growth and osmoregulatory performance and body composition. Diets with equal protein ( 57%) and lipid ( 19%) content (dry weight basis) were supplemented with lipid from either anchovy oil (AO) or CO with AO so that CO comprised 0 (0CO), 11% (11CO), 22% (22CO), 33% (33CO), 43% (43CO) or 54% (54CO) of the dietary lipid content. Triplicate groups of juvenile chinook salmon were fed their prescribed diets for 104 days in freshwater (FW) and 31 days in seawater (SW) after a 4-day transition period. Dietary fatty acid compositions reflected the different proportions of AO and CO in the supplemental lipid. Diet treatment had no effect on fish growth, feed intake, feed efficiency, protein utilization, fish mortality or terminal whole body water and ash percentages. Whole body lipid percentages were higher in 11CO and 43CO fish than in 33CO fish and in 11CO fish versus 22CO fish. Whole body protein percentages were highest in 33CO, 43CO and 54CO fish and lowest in 0CO and 22CO fish. Terminal whole body fatty acid compositions were influenced strongly by the dietary fatty acid compositions. Haematocrit and muscle water percentages were not affected consistently and plasma Na⁺ and Cl⁻ concentrations were unaffected by diet treatment in FW or 24-h seawater challenges during FW residency. Also, diet treatment had no effect on the physiological parameters after SW residency. We conclude that dietary treatment had no effect on fish growth performance under our experimental conditions. Also, the dietary inclusion of CO neither facilitated nor impaired the transfer of chinook salmon parr to seawater. Thus, CO was found to be an excellent and cost-effective source of supplemental dietary lipid for culture of juvenile fall chinook salmon during freshwater residency.     

Effect of dietary protein and lipid source on the growth, survival, condition indices, and body composition of marron, Cherax tenuimanus (Smith)

A trial was conducted in 12 purpose-built, commercial, drainable, earthen ponds to evaluate the effect of fish and plant protein and lipid source on the growth, condition indices, and body composition of marron (Cherax tenuimanus). Juvenile marron (1.3±0.28 S.E. g) at the stocking densities of three per square meter were fed for a period of 1 year with four different formulated isoenergetic practical diets (D₁, D₂, D₃, and D₄). Three of the test diets (D₁, D₂, and D₃) were isonitrogenous whereas the fourth test diet (D₄) was protein-free. Protein and lipid sources in D₁ were from Lupin (Lupinus albus) whereas protein and lipid sources in D₂ and D₃ were from fish meal. Fish oil (3.5%) was added to D₁, D₃, and D₄ whereas sunflower oil was added to D₂ and D₄ in order to make them isoenergetic. The four test diets were randomly allocated to three replicate ponds.

The lack of protein in D₄ did not significantly influence (P>0.05) the mean final weight and specific growth rate of marron. Survival was low in all ponds (13.82–34.66%) but feeding with D₄ resulted in a significantly (P<0.05) higher survival than marron fed with D₁ and D₂. Feeding a diet containing a combination of fish protein and fish oil (D₃) resulted in significantly higher (P<0.05) wet tail muscles-to-body weight ratio than was observed with other diets. Tail muscles protein level of all marron was significantly lower (P<0.05) at the end of the trial than at the beginning. EPA and DHA in hepatopancreas and tail muscles of marron were affected by the four test diets. The inclusion of plant protein in formulated diets had a negative impact on the pond environment due to significantly higher unionised ammonia levels that resulted in lower survival. Juvenile marron fed with a plant protein diet had significantly lower protein levels in their hepatopancreas compared to those fed with diets containing animal protein. Feeding marron with lupin protein source (D₁) and plant oil (D₂) for 1 year did not alter the lipid content of their hepatopancreas. Four test diets had no influence on the fat content of marron hepatopancreas; however, these test diets significantly reduced the protein content of the tail muscles of marron.

Environmental variables, particularly temperature, nitrogen metabolites, and the natural productivity of the ecosystem, greatly influenced the nutritional requirements of the juvenile marron under culture.     

Substitution of Chaetoceros muelleri by Spirulina platensis meal in diets for Litopenaeus schmitti larvae

The nutritional response of Litopenaeus schmitti larvae to substitution of Chaetoceros muelleri by Spirulina platensis meal (SPM) was evaluated. The substitution levels (S) were 0%, 25%, 50%, 75% and 100%, dry weight basis. Final larval length (FL) ranged from 1.98 to 3.16 mm for the different substitution levels. There was a significant relationship between S and FL, described by the following quadratic equation: FL = 2.853 + 0.01598S − 0.000233S². The substitution level (S) yielding maximum FL was 34.2%. Development index (DI) values ranged from 2.84 to 3.93 and were dependent on substitution level. The corresponding equation was DI = 3.799 + 0.00945S − 0.000189S² (P < 0.01). Maximum DI was obtained at 25.0% substitution. Survival was high (82–87%) and no significant differences were found between treatments. Protein digestibility of either microalgae was high, with 92% for SPM and 94% for C. muelleri, with no significant differences between them. The results in this study indicate that an adequate balance of nutrients in relation to the requirements of the species is critical. To simultaneously improve FL and DI, a 30% substitution of C. muelleri by SPM is suggested. This is equivalent to feeding 0.15 mg larvae^− 1 day^− 1 dry weight basis of a 70% C. muelleri/30% SPM diet, representing 0.078 mg protein larvae^− 1 day^− 1, 0.026 mg lipids larvae^− 1 day^− 1 and 2.732 J larvae^− 1 day^− 1.     

Assessing the implications of variability in the digestible protein and energy value of lupin kernel meals when fed to rainbow trout, Oncorhynchus mykiss

A series of studies were designed to examine the degree of variability in the digestibility of protein and energy from lupin (Lupinus angustifolius) kernel meals when fed to rainbow trout (Oncorhynchus mykiss) and the potential implications of this variability. The digestibility of protein and energy from 10 different varieties of lupin kernel meal was assessed where the test ingredient comprised 30% of each test diet. Digesta was collected using faecal stripping techniques. Digestible protein value ranged from 331 to 508 g/kg DM and digestible energy values ranged from 10.6 to 13.3 MJ/kg DM. To examine the implications of variability in digestible protein and energy value, two lupin kernel meals from the extremes of the protein digestibility range (Lupin-1: AD_N ~ 70% and Lupin-2: AD_N ~ 100%) were chosen for assessment in two growth studies. Soybean meal and a reference diet with fishmeal as the only protein source were also included in the study. In the first growth experiment the test ingredients were included at equal concentrations (40%) in protein-limiting diets (350 g protein/kg DM) and fed at either of two ration levels (restricted and satiety). Diets were formulated on a crude basis so as to place the test variable on the variability in digestible protein value of the diets. In the restricted-fed treatments growth of fish fed the reference diet was highest, but not significantly better than lupin-H. Growth of fish fed the lupin-L diet was significantly poorer than both the reference and lupin-H diets, but not poorer than the soybean diet. The growth responses observed from this experiment clearly showed that the differences in feed intake and/or digestible protein value could be demonstrated in terms of significant differences in growth outcomes. In a second growth study high-nutrient dense extruded diets (400 g protein/kg and 23.5 MJ/kg) were prepared with a more practical level of 25% inclusion of the same test materials. Again the diets were formulated on crude basis so as to place the test variable on the variability in digestible protein and energy value of the diets. Growth of fish restrictively fed the lupin-H diet was highest, but not significantly better than the soybean, reference or lupin-L treatments restrictively fed. Growth of fish satietal fed the soybean diet was significantly poorer than the reference and lupin-H diets, but not compared to the lupin-L diet. The growth responses observed from this experiment showed that the differences in digestible protein and energy value could not be demonstrated in terms of significant differences in growth outcomes, and that feed intake variability and excess nutrient supply masked the effect of this variability; particularly at the satietal feed intake levels.     

Effects of chloramphenicol, erythromycin, and furazolidone on growth of Isochrysis galbana and Chaetoceros gracilis

This study focused on determining the effects of antibiotics on microalgae used as food for scallop larvae. Six different dose levels of chloramphenicol, erythromycin, and furazolidone were added to cultures of Isochrysis galbana and Chaetoceros gracilis. An in vivo experiment was subsequently conducted to determine the effect of chloramphenicol and erythromycin on larval survival of the Pacific calico scallop Argopecten ventricosus in tanks and on the population of its associated bacteria. Results showed that growth of I. galbana was not significantly affected by chloramphenicol or erythromycin at the test doses of 0.5, 1.0, 3.0, 6.0, 9.0, and 12.0 mg/l. C. gracilis was significantly sensitive to erythromycin and chloramphenicol at doses higher than 0.5 and 3.0 mg/l, respectively. Furazolidone inhibited the growth of both I. galbana and C. gracilis at all test doses. Results showed that exposure of scallop larvae to a dose of 6 mg/l chloramphenicol or erythromycin did not significantly affect growth of I. galbana, significantly enhanced survival of the scallop larvae, and inhibited the growth of Vibrio spp. in tanks. This study demonstrated the adverse effect of chloramphenicol, erythromycin and furazolidone on I. galbana and C. gracilis microalgae but the positive effect on survival of the scallop larvae, decreasing associated bacterial population.     

Effects of dietary vegetable oils and varying dietary EPA and DHA levels on intestinal lipid accumulations in Atlantic salmon

High dietary content of vegetable oil (VO) has been associated with increased intestinal lipid accumulations in fish. The extent of this in aquacultured Atlantic salmon (Salmo salar L.) and its health effects are not certain. Samples were therefore collected from two separate feeding trials to investigate the effect of high dietary VO on intestinal lipid accumulations in Atlantic salmon. In the first trial, the fish were fed diets high in plant protein and with fish oil or ~80% of the fish oil replaced with either olive oil, rapeseed oil or soybean oil in a land‐based experimental set‐up. The second trial was performed in sea cages under commercial production conditions, and the fish were fed two dietary concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (9.7% or 5.5% EPA + DHA of total fatty acids). Neither dietary VO nor variations in EPA and DHA led to any significant effects on intestinal health or lipid accumulations. There were, however, indications of a delayed lipid transport in the rapeseed oil‐fed fish of the first trial, possibly caused by high dietary ≥18‐carbon fatty acids and low dietary 16:0 fatty acid and cholesterol.