Effect of esterification on the absorption of astaxanthin in rainbow trout, Oncorhynchus mykiss (Walbaum)

Gastrointestinal and serum absorption of astaxanthin was studied in rainbow trout, Oncorhynchus mykiss (Walbaum) (217 ± 2 g) fed diets supplemented with either esterified astaxanthin (from Haematococcus pluvialis) or free astaxanthin (synthetic, as 8% w/w beadlets) at similar levels (50 mg kg^?1). After 56 days of feeding, there was a significant difference (P = 0.0582) between steady‐state serum astaxanthin concentrations for fish fed free (2.0 ± 0.3 μg mL^?1) or esterified astaxanthin (1.3 ± 0.1 μg mL^?1) at the 90% confidence level. However, following ingestion of a single meal supplemented with free or esterified astaxanthin, the rates of astaxanthin absorption into serum were not significantly different (P > 0.1) (0.8 ± 0.2 µg mL^?1 h^?1 and 1.0 ± 0.4 µg mL^?1 h^?1 respectively). In fish fed both free or esterified astaxanthin, higher absorption (P < 0.05) of astaxanthin by the ileal (0.8 ± 0.14 μg g^?1 and 0.9 ± 0.15 μg g^?1 respectively) compared with the posterior (0.2 ± 0.01 μg g^?1 and 0.3 ± 0.14 μg g^?1 respectively) intestine was recorded. This confirmed the role of the anterior intestine in carotenoid absorption. Non‐detectable levels of esters in digesta taken from the hind intestine suggest the anterior intestine is also the primary region for ester hydrolysis.     

Seasonal production of rainbow trout, Oncorhynchus mykiss (Walbaum), in ponds using different feeding practices

Abstract. In temperate regions of North America, the culture of rainbow trout, Oncorhynchus mykiss (Walbaum), in ponds is constrained to the period mid-November to early April by water temperature. Feeding practice may influence production time. The effects of three feeding practices on growth and body composition of rainbow trout raised in ponds during a 142-day winter growth period were investigated. Feeding practices were (1) hand-feeding to satiation, (2) feeding according to a fish size/water temperature chart, and (3) feeding by demand feeder.
Fish fed to satiation consumed 66% more diet than fish fed according to a feeding chart and 163% more than fish fed by demand feeders. Fish fed to satiation had significantly higher (P < 0·05) weight gain, harvest weight, specific growth rate, and protein gain with no significant increase (P > 0·05) in percentage body fat or feed conversion ratio (FCR) compared to the other two treatments. Survival percentage did not differ significantly (P > 0·05) between treatments. These data indicate that by using satiation feeding, marketable size rainbow trout can be produced in temperate region ponds without increases in percentage body fat or FCR.     

Identification and pathogenicity to rainbow trout, Oncorhynchus mykiss (Walbaum), of some aeromonads

Twelve strains of fish pathogenic aeromonads were identified by 16S rRNA sequencing as Aeromonas bestiarum , A. hydrophila , A. hydrophila subsp. dhakensis , A. salmonicida subsp. salmonicida , A. sobria biovar sobria and A. veronii biovar sobria. Following intramuscular injection, A. hydrophila subsp. dhakensis caused dark liquefying, raised furuncle-like lesions in rainbow trout within 48 h. Extracellular products of all cultures contained gelatinase and lecithinase, and most revealed lipase. Congo red absorption and siderophore production was recorded, but not so the suicide phenomenon or slime production. Sodium dodecyl sulphate polyacrylamide gel electrophoresis profile of the outer membrane proteins (OMP) revealed 10–25 bands, of which major bands were seen in the region of 32.5–47.5 and 62–83 kDa. Marked heterogenicity of the OMP and whole cell protein (WCP) profiles within and among the species was observed. Polypeptides of 83–173 kDa were detected in the WCP profile of the cultures, but they were not expressed in OMP fractions.     

The effect of glycyrrhizin on rainbow trout, Oncorhynchus mykiss (Walbaum), leucocyte responses

Abstract. Treatment of rainbow trout macrophages with glycyrrhizin (GL), an aqueous extract of licorice, Glycyrrhiza glabra , enhanced their respiratory burst activity. Maximal effects were seen using concentrations of 10–100 μml⁻¹. GL also modulated trout lymphocytes, increasing proliferation responses to the mitogen phytohaemagglutinin some two-fold over a range of GL concentrations. In addition, GL elicited the release of a macrophage activating factor (MAF) from head kidney leucocytes, as assessed by the ability of generated supernatants to increase respiratory burst activity of target macrophages. MAF activity was most apparent using 100 μg ml⁻¹ GL to induce MAF release and a 48-h incubation period with the target macrophages. Finally, GL was shown to enhance the release of MAF in response to the mitogen concanavalin A. The possible use of GL as a stimulant of fish innate defences is discussed.     

Growth performance and carcass composition of rainbow trout, Oncorhynchus mykiss (Walbaum), fed the (β-agonist ractopamine

Ractopamine is a synthetic catecholamine analogue which is known to improve feed efficiency, reduce carcass fat and increase lean muscle growth in terrestrial livestock (e.g. sheep, pigs, cattle and chickens), and therefore, may be useful in aquaculture production if similar effects can be elicited in fish. In the present study, rainbow trout, Oncorhynchus mykiss (Walbaum), (mean initial weight of 196 g) were fed the β-agonist ractopamine at 0, 5, 10, 20 or 40 parts per million (p.p.m.) of the diet for 12 weeks in a complete, randomized experimental design. Growth and feed intake were monitored throughout the experiment. Carcasses were analysed for protein, fat, energy and ash prior to the initiation of the trial, and subsequently, every 4 weeks. The growth rate, expressed as thermal-unit growth coefficient (TGC) was higher (P ≤ 0.05) in the 10 p.p.m. groups during weeks 1-4 and when averaged over all treatment weeks. Feed efficiency was significantly higher (P ≤ 0.05) in fish fed 10 and 5 p.p.m. ractopamine compared with controls during weeks 1-4 and 4-8, respectively, and was significantly higher (P ≤ 0.05) in all treated groups when averaged over the 12-week treatment period. Compared with controls, carcass protein was elevated in fish fed 5 p.p.m. ractopamine for 4 weeks, and in those offered 10 and 20 p.p.m. for 8 weeks. Carcass fat was significantly lower (P ≤ 0.05) in fish fed 10 p.p.m. ractopamine for 8 weeks, although the 40 p.p.m. group exhibited elevated fat levels in comparison with control fish. The data suggest that rapidly growing rainbow trout demonstrate age-related growth and carcass compositional trends similar to those of mammals, in that feed efficiency and carcass protein decrease while carcass fat increases with age. Oral administration of ractopamine to rainbow trout only modestly altered growth performance, and the trends were not as dramatic nor as consistent as those observed in homeothermic vertebrates fed ractopamine. Out of the concentrations tested, feed incorporation of ractopamine at 10 p.p.m. elicited the most pronounced effect on growth performance.     

The degree of carotenoid esterification influences the absorption of astaxanthin in rainbow trout, Oncorhynchus mykiss (Walbaum)

The absorption of astaxanthin from diets (30 mg kg^?1 inclusion) supplemented with either unesterified astaxanthin; isolated astaxanthin monoesters, diesters or a cell‐free carotenoid extract from Haematococcus pluvialis were studied in rainbow trout (>200 g). No significant differences (P > 0.05) were recorded in the apparent digestibility coefficients (ADC) (≈60–65%) between astaxanthin sources. However, following consumption of a single meal, peak serum astaxanthin levels at 32 h (≈1.0–1.6 μg mL^?1) were significantly higher (P < 0.05) in fish fed unesterified astaxanthin and astaxanthin monoester, compared to fish fed astaxanthin diester and the cell free extract. However, no significant differences (P > 0.05) were recorded in serum astaxanthin uptake rates between sources of astaxanthin. Results suggest that the extent of carotenoid esterification negatively influences the peak serum levels of astaxanthin in rainbow trout.     

Quantification of Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum), tissues by qPCR

A qPCR assay was developed for rapid and sensitive detection of Flavobacterium psychrophilum, the aetiological agent of bacterial cold-water disease and rainbow trout fry syndrome in salmonid fish worldwide. A set of F. psychrophilum-specific primers based on 16S rRNA gene sequences was designed and validated for specific detection and quantification of DNA isolated from representative strains of F. psychrophilum. The qPCR assay exhibited a high specificity for the 16S rRNA gene of F. psychrophilum (from 4 × 10(8) down to 11 copies per reaction) but not for other Flavobacterium species or other bacteria including fish pathogens. This qPCR-based method proved to be useful in the quantification of the F. psychrophilum titre present within organs dissected out from diseased fish. As the F. psychrophilum genome contains six copies of the 16S rRNA gene, we could infer a limit of detection corresponding to two bacteria per reaction, corresponding to 800 bacteria per fish tissue sample, and therefore 20 F. psychrophilum cells mg(-1) of tissue (for sample weighing 40 mg). The qPCR assay reported here could be a useful tool for veterinary diagnostic laboratories to monitor the F. psychrophilum infection level in fish farms.     

Bacterial pathogens in rainbow trout, Oncorhynchus mykiss (Walbaum), reared at Danish freshwater farms

During a 2-year period, bacterial fish pathogens were monitored on five rainbow trout, Oncorhynchus mykiss (Walbaum), freshwater farms in Denmark. A total of 1206 fish were examined and 361 bacterial isolates were identified phenotypically. Enteric redmouth disease, furunculosis and rainbow trout fry syndrome/coldwater disease were recorded. Infections caused by Flavobacterium psychrophilum occurred most frequently, but only one outbreak of enteric redmouth disease caused by Yersinia ruckeri serotype O1 and one of furunculosis caused by Aeromonas salmonicida were recorded during the monitoring period. Flavobacterium psychrophilum was isolated on all farms, both during disease outbreaks and from fish without any signs of disease. Serological investigations of F. psychrophilum showed that serotype Th was the dominant serotype found. The serotypes Th and Fd were involved in disease outbreaks of fry and larger fish. All isolates of F. psychrophilum showed proteolytic activities; however, a few isolates, belonging to serotype FpT did not degrade elastin and were not associated with mortality. Increasing resistance problems to oxytetracycline were demonstrated. More than half of the F. psychrophilum isolates showed resistance to oxolinic acid and oxytetracycline. No antibiotic resistant isolates were found among Y. ruckeri and A. salmonicida .     

Influence of multiple amino acid supplementation on the performance of rainbow trout, Oncorhynchus mykiss (Walbaum), fed soya based diets

Using rainbow trout, Oncorhynchus mykiss (Walbaum), of approximately 50 g, a 63 day feeding trial was undertaken to evaluate the efficacy of individual and multiple amino acid supplements in diets where soyabean meal was used as the principle protein source. Iso-nitrogenous diets (45% crude protein) were formulated where the control diet (F) contained fishmeal as the reference protein and a solvent extracted soyabean meal (S) replaced approximately 66% of this protein source. The soyabean containing diets were then supplemented with crystalline amino acids thus; methionine only, dual supplemented with two methionine and lysine levels and finally, a supplement comprising methionine, lysine, tryptophan, threonine, arginine and histidine. The results showed that soyabean meal (SBM) was inferior to the reference protein when SBM was used to replace 66% of the fishmeal and that no significant restoration in growth, feed efficiency and apparent net protein utilization was obtained by either methionine only or dual methionine and lysine supplementation. However, by comparison with the fish fed the unsupplemented, single and dual supplemented diets, multiple amino acid incorporation was associated with significantly improved percentage weight gain, specific growth rate and marginal improvements in apparent net protein utilization. However, performance was not equal to that of the fish fed the fishmeal based control diet. The results are discussed with respect to the level of each of the essential amino acids (EAA) as a proportion of the total EAAs (A/E index) of test diets by reference to the whole body tissue amino acid profile of rainbow trout.     

Apparent absorption efficiencies of amino acids in rainbow trout, Oncorhynchus mykiss (Walbaum), fed diets containing bacterial single-cell protein

The aim of the present study was to compare the apparent absorption efficiency of individual indispensable and dispensable amino acids in diets containing varying proportions of bacterial single cell protein (BSCP) and fishmeal when fed to rainbow trout, Oncorhynchus mykiss (Walbaum). Four diets were formulated to contain a total of 45.8% crude protein, of which 0% was from BSCP in diet 1 (BSCP-0), 25% in diet 2 (BSCP-25), 62.5% in diet 3 (BSCP-62.5) and 100% in diet 4 (BSCP-100); the remainder of the protein was from fishmeal. The four diets were isonitrogenous and had similar amino acid compositions. Duplicate groups of 25 fish were fed one of the four experimental diets at the rate of 20 g kg^?1 body weight per day for 132 days. Apparent absorption efficiency was measured over a 45-day period during the growth trial. Results from two methods of faeces collection and two indicators of absorption efficiency were compared. The amino acid absorption efficiency of BSCP-100 was significantly lower (P < 0.05) when compared to the other three diets and absorption of some of indispensable amino acids was below the requirement level. The variation seen in individual amino acids absorption efficiencies between diets suggests that this parameter may be important in fish nutritional investigations.     

Gut histology,immunology and the intestinal microbiota of rainbow trout,Oncorhynchus mykiss (Walbaum), fed process variants of soybean meal

Processed soybean meal ingredients have become an emerging plant‐based protein used in aquafeed formulations and have been shown to influence gastrointestinal health and microbiota in cultured species. Two 60‐day feeding trials with juvenile rainbow trout (Oncorhynchus mykiss, Walbaum) were conducted to compare the effects of fishmeal (FMC), defatted soybean meal (SBM), bioprocessed soybean meal (BSBM) and commercial soy protein concentrate (CSPC) ingredients on intestinal histology, innate immunity and microbiota profiles. Results indicated no significant differences in intestinal immunoglobulin concentrations (p = .41) or gut leukocyte phagocytosis at day 15 samplings (p = .41). Intestinal lysozyme content was found to be highest at day 60 in the BSBM treatment group (p < .01). No significant differences across treatments were observed in histological profiles at day 0 (p = .75), day 15 (p = .08) and day 60 (p = .22). ANOSIM analysis for microbiota beta diversity showed differences among all treatment groups, with the exception of the high and low levels of BSBM (p = .40). There were also differences observed in the alpha diversity ratios, with significant differences in gut communities among dietary groups as demonstrated by Chao‐1 (p < .01) and Shannon indices (p = .02). The incorporation of processed soy‐based proteins alters the microbial community composition within the distal intestine, and modulates lysozyme concentrations within the distal intestinal tissues.     

Effect of alternate distribution of astaxanthin on rainbow trout (Oncorhynchus mykiss) muscle pigmentation

The aim of the present study was to determine the effects of the use of astaxanthin alternate feeding on rainbow trout pigmentation in term of astaxanthin serum concentration, muscle colour and astaxanthin muscle retention. Four hundred and fifty rainbow trout were fed the same basal diet supplemented with two different astaxanthin levels, 100, and 200 mg astaxanthin kg^? 1 of diet, hereafter designated as AX100 and AX200, respectively. An additional astaxanthin-free (AX0) diet was used. The experimental treatments were as follows: (1) REF = AX100 diet at each meal each day, served as reference; (2) SD1 = AX100 diet at each meal the first day followed by AX0 diet at each meal every second day; (3) SD2 = AX100 diet and AX0 diet in alternate meals each day; (4) R2 = AX200 diet and AX0 diet in alternate meals each day; (5) R4 = AX200 diet at the first meal the first day followed by AX0 diet at the second meal the first day and at each meal every second day. Fish were fed the experimental feeding schedule for 42 days.At the end of the experiment there were no significant differences among fish fed the different feeding schedules in term of final mean weight, specific growth rate and feed efficiency ratio. SD2 fish group displayed the highest (P < 0.05) astaxanthin serum concentration and the R4 fish group the lowest one. REF and R2 fish groups showed similar astaxanthin serum concentrations. Muscle chroma showed the most pronounced effect. It increased significantly for all fish groups during the experiment. At the end of the experiment REF and R2 fish groups displayed higher values than SD1 and R4 fish groups. Muscle astaxanthin concentrations increased significantly during the experiment whatever the astaxanthin feeding schedule. At the end of the experiment, the highest muscle astaxanthin concentration was recorded for R2 fish group while the lowest was noted for R4 fish group. Except for SD1 and R4 fish groups, muscle astaxanthin retention decreased significantly during the experiment. At the end of the experiment, muscle astaxanthin retention coefficients for SD2 fish group were significantly higher than those for REF fish group. The results reported here provide further evidence of the potential applicability of alternate astaxanthin feeding on rainbow trout pigmentation. Extending the optimisation of the SD2 treatment will therefore be subject for future studies. Its application could result in cost saving in the fish farming industry.     

Immunogenicity of amoebic antigens in rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. Rainbow trout developed a humoral immune response against numerous antigens of sonicated amoebae which were emulsified with Freund's complete adjuvant and injected into the peritoneum. The amoebae were cultured from the gills of Atlantic salmon, Salmo salar L., affected by amoebic gill disease. Antibodies in fish sera were detected by both enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Non-spedfie reactivity in fish serum against Escherichia coli, the bacterium used in co-cultivation of amoebae in vitro, was removed by immunoadsorption. Results obtained using ELISA and immunoblotting were comparable and indicated no significant difference in response to immunization with 10, 50 or 250 μg of sonieatcd amoebic protein. Amoebae contained immunogenie components of > 100, 100, 89, 49, 37 and 34kDa.