Dietary impact of algal and artificial diets, fed at different feeding rations, on the growth and fatty acid composition of Tapes philippinarum (L.) spat

The effect of lipid supplementation and algal ration on growth and fatty acid composition of juvenile clams, Tapes philippinarum was investigated. A diet of Tetraselmis suecica was supplemented with a lipid emulsion and fed at a daily ration of 0.5, 1.0 and 1.5% WW DW⁻¹ day⁻¹. A mixed algal diet of Isochrysis galbana (clone T-Iso) and T. suecica (1:1 on DW basis) and starvation functioned as positive and negative control treatments, respectively. T. suecica, which contains eicosapentaenoic acid (20:5n−3) but no docosahexaenoic acid (22:6n−3), was supplemented with 50% (% of algal dry weight) of a 22:6n−3-rich emulsion. Higher growth rates of T. philippinarum were associated with higher T. suecica feeding rations. Lipid supplementation resulted in significantly better growth rates of T. philippinarum at a T. suecica ration of 0.5, 1.0 and 1.5% WW DW⁻¹ day⁻¹. The growth promoting effect of lipid supplementation was significantly reduced during 4 consecutive weeks or by increasing the algal feeding ration. Increasing T. suecica rations were associated with a decrease in the % composition of 22:6n−3 while the 20:5n−3% composition was hardly affected in the total lipids of T. philippinarum. On the contrary, the absolute concentration of 22:6n−3 (μg per gram dry weight, μg g DW⁻¹) in the total lipids of T. philippinarum was similar at all T. suecica rations while the 20:5n−3 concentration significantly increased. The major effect of lipid supplementation on the fatty acid composition of the clams, was a significant increase of the 22:6n−3% composition in the total lipids, polar lipids and triglycerides. In contrast with 20:5n−3, the effect of lipid supplementation on the absolute 22:6n−3 content (μg g DW⁻¹) was more pronounced at a higher T. suecica ration. After a starvation period of 4 weeks, the concentration (μg g DW⁻¹) of all fatty acids was significantly reduced, except the n−7 monoenoic fatty acids (MUFA) and the non-methylene interrupted dienoic fatty acids (NMID). The concentration (μg g DW⁻¹) of the NMID in starved clams and clams fed different diets was similar to the concentration in the initial sample. This indicated that growing spat was actively synthesising NMID and they were selectively retained in unfed animals. The results indicated that either dietary lipid or 22:6n−3 was a growth-limiting factor when T. suecica was fed at a ration of 0.5, 1 and 1.5% WW DW⁻¹ day⁻¹. A selective retention in starved clams and a preferential accumulation in fed clams was observed for 22:6n−3 but not for 20:5n−3 which indicated a greater importance of 22:6n−3 compared to 20:5n−3.     

Lactic acid fermentation of wheat and barley whole meal flours improves digestibility of nutrients and energy in Atlantic salmon (Salmo salar L.) diets

The effects of lactic acid fermentation of wheat and barley whole meal flours (WMF) on digestibility parameters in Atlantic salmon were studied. The WMFs were inoculated with a specific Lactobacillus strain and fermented for 16 h at 30 °C prior to mixing with other feed ingredients and processing by extrusion. Fermentation of the WMFs significantly decreased total starch (P<0.01) and total mixed-linked (1–3)(1–4)-β-glucan (P<0.001) contents in both cereals. Soluble β-glucans decreased (P<0.001) from 25.2 to 12.0 g kg⁻¹ dry matter in barley WMF during fermentation. In Experiment 1, where diets with 24% untreated wheat or barley WMF and 12%, 24%, or 36% fermented WMFs were fed to Atlantic salmon (0.5 kg) for 25 days, there were indications of improved starch and fat digestibility by fermentation. Experiment 2 comprised diets containing 24% untreated or fermented wheat or barley WMFs fed to each of triplicate groups of Atlantic salmon (0.5 kg) during a 17-day experimental period. This experiment showed that protein (P<0.001), fat (P<0.05) and energy (P<0.001) were more efficiently digested in diets with wheat than in diets with barley. Apparent digestibility of starch was greatly improved by fermentation (P<0.001), more in barley diets (from 47.5% to 67.0%) than in wheat diets (from 51.7% to 65.4%). Improvements in digestibility of fat (P<0.05) and energy (P<0.001) were obtained by fermenting the cereals. The absorption of Na was higher for salmon fed wheat than barley diets (P<0.05). Fermentation resulted in improved Na absorption (P<0.01); from 68.8% to 73.2% for wheat diets and from 60.3% to 71.7% for barley diets. Fermentation caused a significant (P<0.05) improvement in Zn absorption from 32.7% to 40.5% for wheat diets and from 33.2% to 43.5% for barley diets. This may be related to the significant reduction in phytic acid levels seen in both fermented cereals (P<0.001). In conclusion, the potential of wheat and especially barley as ingredients in salmon diets is greatly improved by fermentation.     

Effects of docosahexaenoic, eicosapentaenoic, and arachidonic acids on the early growth, survival, lipid composition and pigmentation of yellowtail flounder (Limanda ferruginea): a live food enrichment experiment

The role of dietary ratios of docosahexaenoic acid (DHA, 22:6n−3), eicosapentaenoic acid (EPA, 20:5n−3) and arachidonic acid (AA, 20:4n−6) on early growth, survival, lipid composition, and pigmentation of yellowtail flounder was studied. Rotifers were enriched with lipid emulsions containing high DHA (43.3% of total fatty acids), DHA+EPA (37.4% and 14.2%, respectively), DHA+AA (36.0% and 8.9%), or a control emulsion containing only olive oil (no DHA, EPA, or AA). Larvae were fed differently enriched rotifers for 4 weeks post-hatch. At week 4, yellowtail larvae fed the high DHA diet were significantly larger (9.7±0.2 mm, P<0.05) and had higher survival (22.1±0.4%), while larvae fed the control diet were significantly smaller (7.3±0.2 mm, P<0.05) and showed lower survival (5.2±1.9%). Larval lipid class and fatty acid profiles differed significantly among treatments with larvae fed high polyunsaturated fatty acid (PUFA) diets having higher relative amounts of triacylglycerols (18–21% of total lipid) than larvae in the control diet (11%). Larval fatty acids reflected dietary levels of DHA, EPA and AA while larvae fed the control diet had reduced amounts of monounsaturated fatty acids (MUFA) and increased levels of PUFA relative to dietary levels. A strong relationship was observed between the DHA/EPA ratio in the diet and larval size (r²=0.75, P=0.005) and survival (r²=0.86, P=0.001). Following metamorphosis, the incidence of malpigmentation was higher in the DHA+AA diet (92%) than in all other treatments (50%). Results suggest that yellowtail larvae require a high level of dietary DHA for maximal growth and survival while diets containing elevated AA exert negative effects on larval pigmentation.     

Amino acid-chelate: a better source of Zn, Mn and Cu for rainbow trout, Oncorhynchus mykiss

This study aimed to evaluate the amino acid-chelated trace elements as dietary supplement to rainbow trout. Three diets were formulated containing trace elements either from the inorganic salt (SF) or amino acid-chelate (AM). Diets 1 (SF) and 2 (AM) contained the same amount of trace elements from inorganic and amino acid-chelates, respectively. Diet 3 (AM-Hf) was added with trace elements from amino acid-chelatex at one-half of their levels in Diets 1 and 2. Each diet was fed for 15 weeks to three groups of 30 fish each, with an average weight of 1.52±0.21 g. Growth of fish was not affected by the treatment (P>0.05). However, bone (P<0.01) and liver (P<0.05) Cu contents were higher in the AM than the SF group. Similarly, hematocrit level (P<0.05) and alkaline phosphatase (ALP) activity (P<0.01) were higher in the chelate-fed fish. Further, DNA polymerase and CuZnSOD expression in the AM group was highly upregulated (P<0.05) compared to the SF fed fish as quantified by RT-PCR. Absorption and whole body retention of Mn and Zn from the AM were higher (P<0.05) than the inorganic salt. Half supplementation of those fed the elements from AM was at par with the full provision from the inorganic source tested.     

The influence of carboxymethylcellulose on growth, digestion, gastric emptying time and body composition of tilapia

Carboxymethylcellulose (CMC) was incorporated into each of two experimental diets at one of four levels (2, 6, 10 and 14%). Each of the eight diets was fed to male tilapia hybrids (Oreochromis niloticus × O. aureus) for 9 weeks. The experiment was conducted in a closed, filtered and recirculated rearing system. The fish were fed the appropriate diets at a rate of 3% of body weight per day, 2 times a day, 6 days a week. The water temperature was maintained at 26°C. Tilapia fed diets containing, 6, 10 and 14% CMC showed significantly poorer (P<0.05) feed conversion rates than fish fed diets containing 2% CMC. The dry matter digestion coefficients of the diets did not differ significantly but generally declined as CMC level increased. Gastric emptying times decreased as the level of inclusion of CMC in the diets increased. Crude protein content and ash content of fish were similar in all dietary groups. Fish fed diets containing 6, 10 and 14% CMC had significantly lower (P<0.05) lipid content than fish fed diets with 2% CMC.     

Water quality control using Spirulina platensis in shrimp culture tanks

A cyanobacterium (Spirulina platensis) was co-cultured with black tiger shrimp (Penaeus monodon) for water quality control. We evaluated the effects of: (1) three S. platensis trial conditions on inorganic nitrogen concentrations at one shrimp density (S. platensis trial conditions included: absent, nonharvested and semicontinuous harvesting) and (2) two shrimp densities on inorganic nitrogen concentrations, with and without S. platensis. Semicontinuous harvesting of S. platensis at one shrimp density resulted in significantly reduced (P<0.05) inorganic nitrogen concentrations (NH₄, NO₂ and NO₃). With S. platensis absent, ammonium and nitrite concentrations ranged from 0.5 to 0.6 mg l⁻¹, while nitrate concentrations ranged from 16 to 18 mg l⁻¹ by day 44. With nonharvested S. platensis, considerable variability occurred with nitrogen concentrations. Semicontinuous harvest of S. platensis reduced nitrate to 4 mg l⁻¹, while ammonium and nitrite ranged from 0.0 to 0.15 mg l⁻¹, respectively. The factorial evaluation of shrimp density versus presence and absence of S. platensis resulted in greatly reduced nitrogenous compounds with S. platensis present regardless of shrimp density, and only moderately increased nitrogen with greater shrimp density. Without S. platensis, all nitrogen compounds were substantially elevated and shrimp survived was significantly reduced at high shrimp density.     

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.     

Dietary folic acid requirement determined for grass shrimp, Penaeus monodon

A feeding trial was conducted to determine the dietary folic acid requirement of juvenile grass shrimp, Penaeus monodon. Purified basal diets were formulated using vitamin-free casein as the protein source. Graded levels (0, 1, 2, 5, 10, 15, 30, and 60 mg folic acid/kg diet) of folic acid were added to the basal diet, resulting in eight dietary treatments in the experiment. Each diet was fed to three replicate groups of shrimp initially averaging 0.79±0.01 g for 8 weeks. Shrimp fed diets supplemented with 2 mg folic acid/kg diet had greater (P<0.05) weight gain and hepatopancreatic folic acid concentrations than shrimp fed the unsupplemented control diet. Feed efficiency (FE) was highest in shrimp fed the diets supplemented with 2 mg folic acid/kg diet, followed by the groups fed 1 mg/kg, then the control group. Hepatosomatic index (HSI) was highest in shrimp fed the control diet, followed by 1 mg folic acid/kg and lowest in shrimp fed diets supplemented with 2 mg folic acid/kg. Weight gain percentage, hepatopancreatic folic acid concentration and hepatosomatic index of the shrimp analyzed by broken-line regression indicated that the adequate dietary folic acid concentration in growing P. monodon is 1.9–2.1 mg/kg diet.     

Deposition of astaxanthin in fillets of Atlantic salmon (Salmo salar) fed diets with herring, capelin, sandeel, or Peruvian high PUFA oils

To elucidate whether absorption and deposition of dietary astaxanthin are influenced by the use of different dietary fish oils having different melting points and fatty acid profiles, triplicate groups of individually labeled Atlantic salmon (initial average weight 569 g) were reared in 2 m² tanks, supplied with saltwater. The fish were fed four different experimental diets coated with either herring oil, capelin oil, sandeel oil, or a Peruvian oil high in polyunsaturated fatty acids. Salmon fed diets containing Peruvian high PUFA oil had significantly higher (13%) fillet carotenoid content than salmon fed herring oil (P<0.05). Astaxanthin retention was significantly higher (P<0.05) in salmon fed capelin and Peruvian high PUFA oil than in the two other groups. The effects of dietary oil supplements on carotenoid deposition were rather small and require verification. Redness of fillets and blood plasma astaxanthin levels did not differ among treatments. Liver weights, body weights, as well as specific growth rates, and feed conversion ratios were unaffected by dietary oil source. Significant (P<0.05) positive linear relationships were found between final fillet idoxanthin concentration and total saturated fatty acids in supplement oils, astaxanthin and total monounsaturated fatty acids, and redness and total n−3 fatty acids, whereas the relationships between redness and total monounsaturated fatty acids and astaxanthin and total n−3 fatty acids were negative.     

Feeding has the largest effect on the ammonia excretion rate of the southern rock lobster, Jasus edwardsii, and the western rock lobster, Panulirus cygnus

The total ammonia nitrogen (TAN) excretion of spiny lobsters Jasus edwardsii and Panulirus cygnus, was determined in relation to temperature, body weight, emersion, daily rhythm and feeding. Temperature and body weight had large influences on the rate of TAN excretion. Exponential relationships were found between temperature (T) and TAN excretion of both species. These were described by the following equations: J. edwardsii Log₁₀ TAN=0.041T−3.57 (r²=0.979, F=143.2, P=0.001), P. cygnus Log₁₀ TAN=0.057T−3.90 (r²=0.987, F=302.2, P<0.001). TAN excretions of both species were positively correlated to body weight (W), and the relationships were described by the following equations: J. edwardsii Log₁₀ TAN=0.473 log₁₀ W−1.704 (r²=0.42, F=14.05, P=0.001), P. cygnus Log₁₀ TAN=0.499 log₁₀ W−1.346 (r²=0.69, F=44.18, P<0.001). TAN excretion increased significantly when lobsters were re-immersed after a 30 min period of emersion. However, it returned to pre-emersion levels by the second hour of re-immersion. Daily rhythm resulted in a significantly higher nocturnal TAN excretion rate for J. edwardsii; no daily rhythm was observed for P. cygnus. Feeding had the largest influence on TAN excretion, with maximum increases of 6.28 (J. edwardsii) and 5.60 (P. cygnus) times the pre-feeding level. TAN excretion rates remained significantly higher than the pre-feeding levels for an extended period (26 h, J. edwardsii; 30 h, P. cygnus). Implications for the use of purging tanks in lobster holding facilities and for the design of biofiltration systems are discussed.     

Dietary phytase supplementation and the utilisation of phosphorus by Atlantic salmon (Salmo salar L.) fed a canola-meal-based diet

The effect of phytase supplementation to a canola-meal-based diet on phosphorus utilisation in Atlantic salmon was studied in a two-by-two factorial design. Diets were prepared without phytase or inorganic phosphorus supplementation, with phytase, with supplemental inorganic phosphorus and with both phytase and supplemental inorganic phosphorus. Available phosphorus was set below requirement and the total phosphorus set to meet requirements for salmonids. After 12 weeks, there were no significant differences in survival, feed intake and weight gain between diets. There was an interaction effect between supplements on bone ash, bone phosphorus and whole-body phosphorus so that adding phytase, inorganic phosphorus, or both resulted in significantly (P<0.05) higher values for these parameters. An interaction effect was also observed for phosphorus digestibility, phosphorus retention efficiency and phosphorus load. Phosphorus digestibility and retention efficiency were significantly (P<0.05) higher, and phosphorus load was significantly (P<0.05) lower in fish fed the phytase supplemented diet compared with diets containing supplemental inorganic phosphorus. In conclusion, phytase increased phosphorus availability, therefore reducing the need to add inorganic phosphorus and reducing phosphorus waste from plant-meal-based diets for Atlantic salmon.     

Protein requirement of jundia fingerlings, Rhamdia quelen, at two dietary energy concentrations

A feeding trial was carried out where five protein concentrations [26%, 29%, 33%, 37%, and 41% crude protein (CP)] were fed to jundia fingerlings at two dietary energy concentrations [3200 and 3650 kcal metabolizable energy (ME)/kg diet] to establish the protein requirement for this species. Triplicate groups of 23 fish (average weight 1.52±0.34 g) were each stocked in 120-l aquaria and fed semipurified diets twice a day to apparent satiation. After 90 days, fish weight gain (WG), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), energy retention (ER), apparent net protein utilization (ANPU), feed consumption (% body weight) and body composition (crude protein, fat, and ash) were affected by diet composition. Weight gain, SGR, FE, ANPU and ER increased (P<0.05) as the dietary protein concentration increased up to 33% and 37%, for fish fed diets containing 3650 and 3200 kcal, respectively. A significant interaction between dietary protein and energy was observed for WG, SGR, and PER. Feed consumption was not affected by dietary energy concentration, but decreased with increasing dietary protein concentration (P<0.05). Body fat decreased as dietary protein increased at both energy concentrations, but was higher in fish fed the 3650 kcal diet. The opposite was observed for body protein, ER, and ANPU (P<0.05). Our findings demonstrate that jundia presents dietary protein sparing effect when the energy concentration increases from 3200 to 3650 kcal/kg. Jundia protein requirement, determined by the broken line method, is between 32.6% and 37.3% CP, depending on dietary energy concentration.     

Beneficial effects of n-3HUFA enriched Artemia as food for larval palmetto bass (Morone saxatilis × M. chrysops)

The beneficial effects of feeding n−3 highly unsaturated fatty acids (HUFA ≥ 20 carbon fatty acids with three or more double bonds) to palmetto bass (striped bass × white bass) larvae, 4–30 days of age, were studied using Artemia diets enriched with six n-3HUFA levels. Dietary n-3HUFA concentrations were < 0.03% (control diet), 0.33%, 0.63%, 0.87%, 1.26%, and 2.27% of dry-wt Artemia. Larval n-3HUFA contents were reduced at a faster rate with decreasing dietary n-3HUFA intake, and were significantly different by 30 days posthatch (4–20 mg g⁻¹ dry-wt fish). Starved larvae selectively conserved endogenous n-3HUFA reserves, indicating an essential role of n-3HUFA in larval development. Mass mortality in the control and 0.33% n-3HUFA diets occurred at metamorphosis (26–28 days posthatch). At harvest, all fish, except those fed the two highest n-3HUFA diets, suffered from handling stress (shock syndrome) with increasing severity (25 to 100%) at decreasing dietary n-3HUFA intake. Recovery from shock syndrome was 100% at the 0.63% and 0.87% n-3HUFA diets, 63% at the 0.33% n-3HUFA diet and 0% at the control diet. Post-harvest survival was similar among the four highest dietary n-3HUFA groups (64–73%), whereas the two lowest n-3HUFA groups differed significantly (0 and 37%). Growth promotion was evident with increased dietary n-3HUFA intake as fish fed the highest n-3HUFA diet were twice the size of those fed the control diet (99 vs 52 mg wet-wt). Final fish sizes at the three highest n-3HUFA diets were similar. Given similar culture conditions, a minimum dietary n-3HUFA intake of 1.26% of dry-wt Artermia is recommended to avoid handling stress and promote growth in larval palmetto bass.     

Dietary vitamin E requirement, fish performance and carcass composition of rohu (Labeo rohita) fry

A 90-day feeding experiment was conducted using flow through system to evaluate the requirement of vitamin E for rohu fry in relation to growth performance and to assess the relationship between the dietary vitamin E levels and carcass composition. Five semi-purified diets supplemented with 0, 50, 100, 150, 200 mg vitamin E/kg dry diet as --tocopheryl acetate were fed to rohu fry (av. wt.±S.E.=0.58±0.01 g) in triplicate groups. At the end of the experiment, growth and dietary performance were evaluated and vitamin E deposition in the tissue, erythrocyte fragility, thiobarbituric acid reactive substances (TBARs) were analyzed. The average net weight gains were 6.82±0.09, 7.38±0.03, 8.20±0.03, 8.07±0.09 and 8.12±0.08 (g/90 days), respectively, for fish fed diets 0, 50, 100, 150, 200 mg vitamin E/kg. The fish fed diets containing less than 100 mg supplemental vitamin E/kg had significantly (P<0.05) reduced weight gain, feed efficiency and other nutritional indices compared to those fed diets supplemented with vitamin E at 100–200 mg/kg. With the increasing level of vitamin E, both TBARs and erythrocyte fragility values were reduced. Regression analysis of weight gain data using broken-line model indicated a minimum vitamin E requirement of 131.91 mg/kg dry diet. The erythrocyte fragility and TBARs were maximum in fish fed the vitamin E-deficient diet. From the present experiment, it may be concluded that the vitamin E requirement of Labeo rohita fry for optimum growth and other dietary performances is 131.91 mg/kg of dry diet.     

Influence of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder, Paralichthys olivaceus

A feeding trial using five semi-purified diets (50% crude protein) was conducted to investigate the effects of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder. The control diet contained casein and gelatin as intact protein sources and four other diets contained 30% casein–gelatin (2:1, w/w) and 20% crystalline amino acids (CAA). CAA were added to the diets to simulate the amino acid pattern found in red sea bream egg protein (REP), Japanese flounder larvae whole body protein (FLP), Japanese flounder juvenile whole body protein (FJP), and brown fish meal protein (BFP), respectively. The test diets were fed to triplicate groups of juveniles (2.75±0.05 g) twice a day for 40 days to evaluate weight gain, survival, feed conversion efficiency (FCE), protein efficiency ratio (PER), and apparent protein utilization (APU). The apparent retention of total dietary amino acids in the whole body and A/E ratios of the whole body were also evaluated. The highest weight gain was observed in fish fed the diet containing the dietary amino acid pattern of BFP followed by fish fed the control, FJP, FLP and the REP diets. Percent survival, FCE, PER and APU were also significantly (P<0.05) affected by the amino acid pattern in the diets, indicating the highest value in fish fed the BFP diet. Except for a few amino acids, the amino acid composition of the whole body did not show marked differences with different dietary amino acid pattern. Results suggest that BFP could be more suitable as a reference amino acid pattern in the diet of juvenile Japanese flounder compared to the amino acid pattern of FLP, FJP or REP.     

Safety and efficacy of emamectin benzoate administered in-feed to Atlantic salmon, Salmo salar L., smolts in freshwater, as a preventative treatment against infestations of sea lice, Lepeophtheirus salmonis (Krøyer)

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg⁻¹ fish day⁻¹ for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg⁻¹ day⁻¹, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg⁻¹ day⁻¹. Among smolts that received 272 μg kg⁻¹ day⁻¹, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified.

In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg⁻¹ day⁻¹ (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P<0.001) on treated fish than on controls. When fish challenged at day 109 were sampled at day 128, efficacy was 44.3%, but survival of chalimus to adult lice on treated fish was lower, and at day 159, efficacy had increased to 73%. These results demonstrate that treatment of salmon smolts with emamectin benzoate in freshwater was well tolerated and highly effective in preventing sea lice infestation following transfer of fish to seawater.     

Studies on aneuploid pearl oyster (Pinctada martensii Dunker) produced by crossing triploid females and a diploid male following the inhibition of PB1

Viable aneuploid embryos of pearl oyster, Pinctada martensii Dunker, were produced by inhibiting the first polar body (PB1) with cytochalasin B treatment in eggs from triploids fertilized with haploid sperm. During the period of growth measurement, aneuploid showed the slowest growth compared with diploid and triploid groups. The body size and weight measurement data showed that there were no differences between aneuploids (as a group) and diploids in body size and weight (P>0.10), but that they were significantly different from triploids (P<0.01). There were no differences between aneuploids within diploid condition (2n±n) and diploids in SL (P>0.1), but significant differences in BW were found (P<0.05). Aneuploids within triploid condition (3n±n) were significantly smaller than triploids in BW (P<0.05), but not different from triploids in SL (P>0.05), and almost identical to diploids in both (P>0.1). These dada indicated that some aneuploids might be associated with growth retardation. Karyotype analysis revealed that there were metacentric, submetacentric or subtelocentric chromosomes losses or gains; aneuploid pearl oysters with the same chromosome numbers had different chromosome composition. Aneuploids are valuable research materials for genetic analysis.     

Yields of cultured Pacific oysters Crassostrea gigas Thunberg improved after one generation of selection

Parental families (G₀) in three lines of Pacific oysters were selected based on live weight and meat yields at harvest. The average live weight yield of progeny (G₁) from crossing G₀-selected lines in seven trials was 9.5% greater than that of nonselected control families and live weight yields were significantly greater (ANOVA, P<0.001) in four out of seven trials. The response to selection was greater if G₁ families were tested at the same site as their parents' selection site rather than at a different site, although this effect was only significant for G₁ families of cohort 5 (P<0.01) but not cohort 7 (P>0.05). A significant genotype×environment interaction affected yields in both cohort 5 and cohort 7 (ANOVA; P<0.001). In addition, correlation between the yields of the same families planted at both intertidal and subtidal sites was positive but weak (cohort 5, r=0.30; cohort 7, r=0.35), indicating that selection for high yield in one environment would likely result in a low correlated response in a different environment. Nonetheless, it was possible to identify six families in cohort 5 and four families in cohort 7 that were among the top 10 families at both sites. Further evaluation of families across a wider range of environments is needed to determine if it is possible to substantially improve yields by selecting “generalist” families that perform well along the whole Pacific coast, or whether it will be necessary to select lines that are suited to particular sites.     

Effects of dietary protein to energy ratios on growth and body composition of juvenile Japanese seabass, Lateolabrax japonicus

A growth experiment was conducted to determine the optimal dietary protein to energy (P/E) ratio for juvenile Japanese seabass (initial average weight 6.26±0.10 g). Nine practical diets were formulated to contain three protein levels (36%, 41% and 46%), each with three lipid levels (8%, 12% and 16%), in order to produce a range of P/E ratios (from 19.8 to 28.6 mg protein kJ⁻¹). Each diet was randomly assigned to triplicate groups of 60 fish in sea floating cages (1.5×1.5×2.0 m). Fish were fed twice daily (06:30 and 16:30) to apparent satiation for 8 weeks. The water temperature fluctuated from 26.5 to 32.5 °C, the salinity from 32‰ to 36‰ and dissolved oxygen content was approximately 7 mg l⁻¹ during the experimental period. The results showed that the growth was significantly affected by dietary P/E ratio (P<0.05). Fish fed the diets with 46% protein (12% and 16% lipid, P/E ratio of 26.7 and 25.7 mg protein kJ⁻¹) had the highest specific growth rates (SGR) (4.26 and 4.24% day⁻¹). However, fish fed the diet with 41% protein and 12% lipid showed comparable growth (4.20% day⁻¹), and had higher protein efficiency ratio (PER), protein productive value (PPV) and energy retention than other groups (P<0.05). No significant differences in survival were found among dietary treatments. Carcass lipid content positively correlated with dietary lipid level irrespective of protein level and inversely correlated with carcass moisture content. Carcass protein and ash contents increased with increasing dietary lipid at each protein level. These results suggest that the diet containing 41% protein and 12% lipid with P/E of 25.9 mg protein kJ⁻¹ is optimal for Japanese seabass.     

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.