The Norwegian seaweed industry is expanding and there is a need for accurate estimates of protein content of seaweed species from Norwegian waters. A solid method to calculate protein content is ...through the sum of the proteomic amino acids; however, it can be expensive and beyond the capacities of many laboratories. The most commonly used method to quantify protein is based on the assessment of crude protein from overall nitrogen content, using the traditional nitrogen-to-protein conversion factor of 6.25. However, this approach can be inaccurate when applied to seaweeds, often resulting in an overestimation of their protein content. Specific nitrogen-to-protein conversion factors, calculated from amino acid composition and total nitrogen, give a more reliable protein quantification in seaweeds. However, no such factors are available for species from Norwegian waters. This study was designed to characterize the amino acid composition of 21 seaweed species from Norwegian waters and use the amino acid data to estimate protein contents of the seaweeds. Crude protein analysis (nitrogen × 6.25) was performed and resulted in overestimation (18–44 %) of the protein content compared to the sum of proteomic amino acids. Specific nitrogen-to-protein conversion factors, calculated for each species, ranged from 3.53 ± 0.1 to 5.13 ± 0.1. This study provides nutritional data on Norwegian seaweeds, covering a relatively wide range of species. Moreover, it is the first study to assess nitrogen-to-protein conversion factors on such species.
Methionine is an indispensable amino acid with an important role as the main methyl donor in cellular metabolism for both fish and mammals. Metabolization of methionine to the methyl donor ...S-adenosylmethionine (SAM) has consequence for polyamine, carnitine, phospholipid, and creatine synthesis as well as epigenetic modifications such as DNA- and histone tail methylation. Methionine can also be converted to cysteine and contributes as a precursor for taurine and glutathione synthesis. Moreover, methionine is the start codon for every protein being synthetized and thereby serves an important role in initiating translation. Modern salmon feed is dominated by plant ingredients containing less taurine, carnitine, and creatine than animal-based ingredients. This shift results in competition for SAM due to an increasing need to endogenously synthesize associated metabolites. The availability of methionine has profound implications for various metabolic pathways including allosteric regulation. This necessitates a higher nutritional need to meet the requirement as a methyl donor, surpassing the quantities for protein synthesis and growth. This comprehensive review provides an overview of the key metabolic pathways in which methionine plays a central role as methyl donor and unfolds the implications for methylation capacity, metabolism, and overall health particularly emphasizing the development of fatty liver, oxidation, and inflammation when methionine abundance is insufficient focusing on nutrition for Atlantic salmon (Salmo salar).
A 10‐week growth trial was run to evaluate effects of myo‐inositol (MI) on growth performance, haematological parameters, antioxidative capacity and salinity stress tolerance of Litopenaeus vannamei. ...Six practical diets supplemented with graded levels of MI (designated as MI0, MI600, MI1200, MI2400, MI 3600 and MI4800 for 448.8, 974.2, 1568.0, 2810.6, 3835.5 and 4893.6 mg/kg diet, respectively) were fed to six replicate groups of L. vannamei (mean initial body weight 0.63 ± 0.00 g). The results showed that significant increment of growth performance was observed in shrimp fed MI600 diet than those fed MI1200 diet. Lipid concentration in whole body of the shrimp fed MI600 diet was significantly increased. Shrimp fed MI0 diet had lower total protein (TP) as compared to shrimp fed the MI‐supplemented diets (except MI4800 diet). In general, lower activities of antioxidant enzymes and higher malondialdehyde (MDA) content in haemolymph and hepatopancreas were recorded in shrimp fed MI0 diet, compared to those fed the MI‐supplemented diets. Reduced survival after 7‐h salinity stress was present in shrimp fed MI0 diet as compared to those fed MI4800 diet. Dietary MI requirement for glutathione peroxidase activity of L. vannamei was 2705 mg/kg diet.
The present study is focused to elucidate the main characteristics of the digestive function of this carnivorous fast-growing fish living at high temperatures. With this aim, we have examined the ...effects of an increased temperature from 30 to 34 °C on the daily pattern of gastrointestinal pH, enzymatic proteolytic digestive activity and the feed transit time in early juveniles of cobia (Rachycentron canadum), a species living in tropical and subtropical waters with an increasing aquaculture production. Fish were fed two meals a day. Gastric luminal pH was permanently acidic (mean pH values: 2.76–4.74) while the intestinal pH increased from neutral/slightly acidic to slightly alkaline when the digesta was present, with an increasing alkalinity from proximal to distal intestine (mean pH values: 6.05 to 7.69). The temperature did not affect the gastric pH but a slightly higher acidity was induced in the intestine at 34 °C.
Pepsin activity showed a daily rhythm at 30 °C with maximum in the middle of the light period, while at 34 °C some hourly changes coinciding with feed adding without a clear daily trend during the 24-h period were observed. The trypsin activity exhibited a daily rhythm at both temperatures with an increase after morning feeding to reach a maximum several hours later. Average pepsin activity during the daily cycle was slightly higher at 34 °C (6.1 and 7.3 U mg−1 BW at 30 and 34 °C respectively), but values were significantly different only at 8 and 24 h after the morning meal. Similarly, the trypsin activity was significantly affected by the temperature only at 8 and 16 h after the morning meal, but daily activity averages were similar (1.20 and 1.29 U g−1 BW at 30 and 34 °C respectively).
The partial transit rates of the first meal in the stomach for each period inter-samplings were higher during the first 4-h period and decreased progressively along the rest of the 24-h cycle at both temperatures, but no significant differences were detected at 30 °C. In addition, the transit was notably faster at 34 °C particularly during the first 8 h after feeding, with rates between 100 and 65% of total volume displaced (intake or released) during each 4-h period. In the intestine the transit rate was relatively constant and similar at both temperatures during 12 h after feeding. Then the rates remained very low during the following 12 h.
Residence time of the first meal was longer at 30 than at 34 °C, particularly in the stomach (12 h:02 min vs 4 h:54 min respectively). In the intestine the difference was not so large (8 h:18 min vs 6 h:24 min respectively). In a parallel study under same conditions, cobia reared at 30 °C grew faster and showed a more favorable feed conversion ratio than those at elevated temperature (34 °C). The present results indicate that at 34 °C, a subtle increase of proteolytic activity cannot compensate for the faster gut transit rate. Therefore, 30 °C is more appropriate temperature for the early on-growing of cobia because at higher temperatures the digestion efficiency decrease being one of the causes for a lower growth.
•Cobia exhibits a permanent gastric acidification.•Water temperature (30 and 34 °C) does not substantially affect the digestive proteolytic activities.•Both stomach and intestine are filled almost simultaneously.•Transit time was much faster and the residence time lower at 34 °C than at 30 °C.
The effects of short-time fasting on appetite, growth, and nutrient were studied in Atlantic salmon (Salmo salar) smolts. Feed deprivation did change the energy metabolism with reduced plasma protein ...and muscle indispensible amino acid levels. Plasma levels of ghrelin were significantly higher in starved salmon compared with fed fish after 2 days, but no differences in circulating ghrelin were found between treatments after 14 days. Two mRNA sequences for ghrelin-1 and ghrelin-2, 430 and 533 bp long, respectively, were detected. In addition, the growth hormone secretagogues-receptor like receptor (GHSR-LR) 1a and 1b were identified. Ghrelin-1 but not ghrelin-2 mRNA levels were affected by starvation in the stomach. Lower ghrelin-1 mRNA levels were detected at day 2 in starved fish compared with fed fish. The mRNA levels of GHSR-LR1a were not affected by starvation. Fasting reduced the phenotypic growth and the transcription of insulin-like growth factor (IGF)-II together with IGF-IIR, but IGF-I mRNA were not regulated in fasted salmon after 14 days. Three IGF-binding proteins (IGFBP) at 23, 32, and 43 kDa were found in salmon, and circulating 23 kDa was significantly increased after 14 days of starvation compared with fed fish, indicating increased catabolism. The levels of IGFBP-1 mRNA were significantly higher in fed and starved fish after 14 days compared to those at the start of the experiment, but no significant difference was observed between the treatments. In conclusion, we have shown that circulating ghrelin and ghrelin-1 mRNA is related to changes in energy metabolism in Atlantic salmon.
The aim of the present study was to combine maximum replacement of fish meal and fish oil with plant ingredients in feed for Atlantic salmon, in order to gain a sound and sustainable net fish protein ...production. The design implied that all known nutrient requirements should be met. Atlantic salmon smolts with an initial weight of 0.3 kg were fed in triplicate either a fully marine control diet or one of three plant based diets through the seawater production phase for 12 months, until final weight of approximately 4 kg. In a maximum plant based diet, 80% of the fish meal was replaced with a mixture of plant protein ingredients and krill meal, while 70% the fish oil was replaced with a mixture of vegetable oils. Two intermediate replacement diets contained either one half of this fish meal replacement level and maximum fish oil replacement, or one half replacement level of fish oil and maximum fish meal replacement. Fish performance was assessed by measuring mortality, feed intake, growth, nutrient digestibility and nutrient utilisation. Specific growth rate was significantly lower in the combined high replacement group compared to the other experimental groups, both for the first 3-month period (12%) and for the complete 12 months (9%) of feeding. The final fish weights were 17% lower in the combined high replacement group and 9% lower in the high plant protein and intermediate vegetable oil group, compared to the marine control and the intermediate plant protein group. Significantly reduced feed intake during the first period and slightly reduced digestibility of 16:0 and starch were identified as possible causes for growth depression, since minor differences in protein or lipid digestibility, feed conversion ratio, and protein and lipid retention were observed.
The maximum fish meal and fish oil replacement represented a net production of fish protein, with 2 kg salmon protein produced per kg fish meal protein fed. This being four-fold more efficient usage of fish meal in the 80% plant protein diets compared to the 100% fish raw material diet.
The aim of the current study was to investigate how freshwater diets impact on immunity in Atlantic salmon smolts in freshwater, during transfer to seawater and in post smolts during the seawater ...stage with and without pancreas disease (PD) infection. Three specific freshwater diets were prepared: (i) A diet similar in composition to commercial salmon freshwater diets (Standard diet); (ii) A diet composed of vegetable oils (rapeseed, palm and linseed oils) mimicking the fat composition in aquatic insects – the natural diet of wild salmon in freshwater (Fatty acid diet); (iii) A diet enriched with possible immune modulating amino acids including dl-methionine, l-lysine, l-threonine and taurine (Amino acid diet). After seawater transfer, all fish were fed the same commercial diet. Head kidneys were extracted, and their leukocytes isolated from smolts right before transfer to seawater, from post smolts one and six weeks after transfer to seawater, and from post smolts in seawater after 8 weeks of ongoing PD infection. In addition, to provoke bacterial or virus induced inflammation in vitro, the individual leukocyte suspension from all fish were stimulated by lipopolysaccharide (LPS) or polyinosinic acid: polycytidylic acid (PIC).
The transfer of smolts from fresh-to seawater changed the transcription of several types of genes. Particularly in isolates from fish fed the Standard or Fatty acid diet in freshwater, overall gene transcription (IL-1β, CD83, INF-γ, cox2, cd36, MGAT2, catalase) declined. However, the Amino acid diet stimulated the LPS induced gene transcription of IL-1β, CD83, Cox2, and INF-γ at this stage. In freshwater smolts, PIC stimulated leukocytes showed higher transcription level of Mx and viperin in the Fatty acid and Amino acid diet groups compared to the Standard diet group. In seawater post smolts, Mx and viperin responded similarly to PIC challenge in all diet groups. Furthermore, leukocytes isolated from PD infected fish, continued responding to PIC, regardless of freshwater diet.
•Modified Amino- and Fatty acid composition in freshwater diets can increase the immunological robustness of salmon parr towards virus infections during the freshwater phase.•The Amino acid composition in freshwater diets can be tailored to be protective against bacterial and oxidative damage in post smolts newly transferred to seawater.•When challenged with PIC, PD virus or PD virus + PIC, salmon immune cells use their energy to produce antiviral molecules, regardless of diet composition in freshwater.
This study sought to investigate whether a “natural diet” (mimicking the fatty acid composition of freshwater aquatic insects eaten by salmon parr) during the freshwater (FW) life stage of pre-smolt ...Atlantic salmon (Salmo salar L.) affected red blood cells and gill fatty acid composition as well as eicosanoid metabolism in gill during smolting at different temperatures. Before being transferred to seawater (SW), salmon parr were fed with a modified (MO) diet containing vegetable oils (rapeseed, palm, and linseed oils) supplemented with eicosapentaenoic acid (EPA) and arachidonic acid (ARA) to completely replace the fish oil (FO). Fatty acid composition in red blood cells and gill tissues was determined before SW transfer and six weeks after. Additionally, the expression of genes associated with eicosanoid metabolism and Na+/K+-ATPase (NKA) activity in salmon gill was examined at different temperatures before SW transfer and 24 h after. The results showed the changes in fatty acid composition, including sum monounsaturated fatty acids (MUFAs), docosahexaenoic acid (DHA), ARA, EPA, and sum n-6 polyunsaturated fatty acids (n-6 PUFA) in both red blood cells and gill tissues at the FW stage were consistent with the fatty acid profiles of the supplied MO and FO fish diets; however sum EPA and DHA composition exhibited opposite trends to those of the FO diet. The proportion of ARA, EPA, and n-6 PUFA increased, whereas sum MUFAs and DHA decreased in the red blood cells and gill tissues of MO-fed fish compared to those fed with the FO diet at FW stage. Additionally, 5-lipoxygenase-activating protein (Flap) expression was downregulated in MO-fed fish prior to SW transfer. During the process of SW transfer at different temperatures, the MO diet remarkably suppressed NKAα1a expression in MO-fed fish both at 12 and 16 °C. The MO diet also upregulated phospholipase A2 group IV (PLA2g4) expression in gills at 8, 12, and 16 °C, but suppressed phospholipase A2 group VI (PLA2g6) expression in gills at 12 °C compared to FO-fed fish at 12 °C and MO-fed fish at 8 °C. The MO diet also upregulated Cyclooxygenase 2 (Cox-2) expression at 8 °C compared to FO-fed fish and increased Arachidonate 5-lipoxygenase (5-Lox) expression in MO-fed fish at 16 °C compared to both FO-fed fish at 16 °C and MO-fed fish at 8 °C. Our study also determined that both SW transfer water temperatures and diets during the FW period jointly influenced the mRNA expression of PLA2g4, PLA2g6, and Lpl, whereas 5-Lox was more sensitive to dietary changes. In conclusion, the MO diet affected the fatty acid composition in gill and in red blood cells. When transferred to SW, dietary ARA supplementation could promote the bioavailability for eicosanoid synthesis in gill mainly via PLA2g4 activation, and potentially inhibit the stress and inflammatory response caused by different water temperatures through dietary EPA supplementation.
•Salmon parr were fed with a MO diet only containing vegetable oils supplemented with EPA and ARA.•The MO diet affected fatty acid composition synergistically in gills and red blood cells at FW stage.•A lasting impact of MO diet during FW stage on fatty acid composition after SW transfer.•The MO diet promoted the eicosanoid biosynthesis in gill during SW transfer.•The MO diet inhibited the inflammatory response during smoltification at different temperatures.
This study aimed to determine the impact of elevated temperature combined with different levels of dietary methionine concentrations on feed intake (FI) and brain expression of selected neuropeptides ...and one receptor involved in appetite control in juvenile cobia (approximately 3.7 g body weight). The genes studies were neuropeptide y,
npy;
agouti-related protein,
agrp
; cocaine- and amphetamine-regulated transcript,
cart;
cholecystokinin,
cck
and melanocortin 4 receptor;
mc4r
. The cobia were reared at typical sea water temperature in Vietnam (30 °C) and elevated temperature (34°C; selected as one of the predicted scenarios of climate change). The fish were fed diets with different levels of methionine: deficient (M9; 9.1 g/kg), sufficient (M12; 12.8 g/kg) and surplus (M16, 16.8 g/kg) for 6 weeks (triplicate tanks per treatment). Both dietary methionine concentration and temperature affected FI in cobia. Dietary methionine deficiency (i.e., M9) and elevated temperature reduced FI in cobia. Temperature significantly influenced the mRNA expression of
agrp
,
cart
,
cck
and
mc4r
. Expression of the orexigenic neuropeptide
npy
was consistently higher before the meal than after the meal for all diets and at both temperatures. At 30°C, prefeeding levels of
npy
correlated with both increased methionine levels and FI. The interaction between dietary methionine and temperature on the levels of brain
npy
expression was significant (P<0.05). There was higher brain expression of
agrp, cart
and
cck
in cobia at 34°C than in fish at 30°C, which was correlated with a lower FI. In conclusion, both feeding, temperature and/or dietary methionine levels affected the brain expression of
npy
and
agrp, cart, cck
and
mc4r.
This suggests that these neuropeptides as well as the mc4r receptor are actively involved in adjusting feed intake to compensate for changing energetic demands, as well as metabolic adjustments due to the variable availability of methionine at elevated temperature.
We previously reported that juvenile Atlantic salmon with mean initial BW 11.5 g offed a methionine deficient diet had lower weight gain due to a reduced protein accretion, while lipid gain was ...unaffected. Muscle of the fish fed the methionine deficient diet was depleted for sulphur amino acids, while in liver, the concentration of these metabolites was maintained within narrow limits. We speculated whether this could be due to an increased muscle proteolysis to support a prioritized liver metabolism in fish fed the low methionine diets. In this study, we assessed whether genes associated with muscle proteolysis increased under methionine deficiency. The composition of the diets was similar to those used previously containing 1.6 or 2.1 g Met/16 g N. We confirmed that the fish fed the low methionine diet gained less protein compared to fish fed the DL‐methionine enriched diet (P = 0.014), but growth did not reduce significantly. Also the deficient fish maintained the concentrations of liver sulphur amino acids and reduced muscle free methionine. Several of the other free amino acids within muscle increased. Further, methylation capacity was maintained in liver but reduced in the muscle (P = 0.78 and 0.04, respectively). Gene expression of muscle IGF‐1 was lower (P = 0.008) and myosin light chain 2 tended (MLC2, P = 0.06) to be reduced in fish fed low methionine diet, concurrently the activity of cathepsins B+L increased (P = 0.047) in muscle of fish fed the low methionine diet. Gene expression of the muscle‐specific E3 ubiquitine ligases (Murf and MaFbx) was not affected by treatment. Thus, the lower protein gain observed in fish fed the low methionine diet may be caused by reduced protein synthesis in line with the reduced IGF‐1 gene expression in the white trunk muscle. Thus, to support metabolism, the dietary protein needs to be balanced in amino acids to support metabolism in all compartments of the body and secure maximal protein gain.
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