Effect of fish feed processing conditions on digestive protease activities, free amino acid pools, feed conversion efficiency and growth in Atlantic salmon (Salmo salar L.)

The responses of the digestive proteases trypsin and chymotrypsin and protein metabolism to differences in feed protein quality were investigated in Atlantic salmon (Salmo salar L.). Two sets of experimental feeds were produced. Each set of high and low quality feeds was provided to either 150 g or 2 kg salmon. Protein in the high quality feeds had significantly higher percentages of free (reactive) sulphydryl (SH) groups than the corresponding feeds based on low quality meals. After 90 days feeding, groups given high and low quality feeds did not differ in their specific growth rates (SGR) in either experiment. However, feed conversion efficiency (FCE) was significantly different between the high and low quality feed groups in 2 kg salmon, where the difference between the high and low feed protein qualities was larger, 10% versus 4% SH/[SH + (S–S)] in 150 g salmon. Higher FCE was preceded by significantly higher trypsin and chymotrypsin specific activities on day 60. SGR, in general, changed after the first month and was stable during the last 2 months in both experiments. Concurrently, both trypsin (T) and chymotrypsin (C) decreased with an increased activity ratio of trypsin to chymotrypsin (T/C ratio), and resulting in significantly lower T/C ratio on day 90 in salmon feeding on high quality feeds in both sizes of fish. Differences in FCE were associated with significant differences in levels of total free amino acids (TFAA) in the plasma and the white muscle, as well as in the ratio of essential to non‐essential free amino acids (EAA/NEAA ratio), free hydroxyproline, and RNA in the white muscle. Interestingly, after 3 days starvation (day 93), 5–7 h postprandial EAA/NEAA ratio in the plasma was significantly lower in the high quality diet groups in both experiments. Trypsin specific activity inversely correlated with muscle TFAA levels in 2 kg salmon, concurrent with higher muscle levels of RNA, lower free hydroxyproline and higher FCE in fish fed higher quality diets.     

Measures of egg quality in induced spawns of the Asian sea bass, Lates calcarifer Bloch

The egg morphometry and lipid and protein components were determined in induced spawns (n=14) of the sea bass, Lates calcarifer, to identify measures of egg quality. Based on fertilization and hatching rates, the spawns were classified either in Group I, (zero fertilization) or Group II (where fertilization and hatching occurred). The egg morphometry did not differ between the two groups (p > 0.05). The total lipid was higher in Group II than in Group I, although the difference was not significant (p > 0.05). The EPA and linoleic acid were significantly higher in Group II (p < 0.05). There were positive correlations between the total saturated fatty acids and fertilization rate (p < 0.05; r=0.58), the total saturated fatty acids and percentage of normal zygotes (p < 0.02; r=0.62), and DHA and the percentage of normal zygotes (p < 0.04; r=0.56). The total protein and FAAs were higher in Group I than in Group II, but the differences were not significant (p > 0.05). Proline, glycine, p-ethanolamine, and aspartic acid were significantly higher in Group II (p < 0.05), while tyrosine and glutamic acid were significantly higher in Group I (p < 0.05). Phosphoserine and fertilization rate were positively correlated (p < 0.03; r=0.60), as well as aspartic acid and hatching rate (p < 0.05; r=0.54). Arginine was negatively correlated with fertilization rate (p < 0.03; r=–0.61) and the percentage of normal zygotes (p < 0.03; r=–0.63). Serine was inversely correlated with yolk volume of the newly-hatched larvae (p < 0.03; r=–0.77). The moisture content of the eggs, which was significantly higher in Group II than in Group I (p < 0.03), was directly correlated with the FAAs:protein ratio (p < 0.03; r=0.76). The present results reveal egg components that may be used as quality measures in induced spawns of sea bass, a euryhaline teleost that spawn pelagic eggs containing an oil globule.     

盐度对凡纳滨对虾肌肉中游离氨基酸含量的影响

通过逐渐升高盐度的方法,分别测定盐度为0、10、20、30和40时凡纳滨对虾肌肉中游离氨基酸(FAA)的含量,实验周期40 h。结果表明:在盐度0~40时,总游离氨基酸(TOFAA)、必需氨基酸(EAA)、非必需氨基酸(NEAA)和鲜味氨基酸(TAA)的含量都随盐度的增加而明显升高(P0.05);特别是当盐度从20增加到30时,肌肉中TOFAA、EAA、NEAA和TAA含量的增高幅度最明显。在凡纳滨对虾肌肉FAA中,含量最高的是甘氨酸(Gly),其次是脯氨酸(Pro)、精氨酸(Arg)和丙氨酸(Ala)。在盐度为0~40范围内,肌肉中主要的渗透压调节因子是甘氨酸(Gly)、脯氨酸(Pro)、精氨酸(Arg)、丙氨酸(Ala)和牛磺酸(Tau);而天冬氨酸(Asp)、丝氨酸(Ser)、缬氨酸(Val)、甲硫氨酸(Met)、异亮氨酸(Ile)、亮氨酸(Leu)和酪氨酸(Tyr)的含量在各盐度组未呈现显著变化(P0.05)。在淡水养殖凡纳滨对虾出售前的40 h,通过提高水体盐度来提高凡纳滨对虾肉质的鲜味是完全可行的。     

Digestive efficiency,free amino acid pools and quality of growth performance in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) affected by light regimes and vaccine types

This study comprised the results of three different seawater trials using unique combination of techniques to study protease digestive efficiency and growth performance quality to illustrate the effects of light regimes and vaccine types in Atlantic salmon (Salmo salar L.). Fish with higher growth had higher trypsin (T) and chymotrypsin (C) specific activities with higher T/C ratio or slope T/C ratio [calculated from the regression between trypsin (y) and chymotrypsin (x) specific activities] in the pyloric caeca. The T/C ratios indicated fish growth rates over a period of 1–2 months, while the slope T/C ratios indicated fish growth rates at sampling. Adaptation period for adjustment to the new environment of continuous light was 70 days, indicated by the differences in trypsin specific activities and the crossing of slope T/C ratio regressions following with the changes in growth rate directions between the control and the treated group. Vaccine types affected fish vertebral growth, and additional continuous light enhanced the impact of vaccines on fish growth during springtime, indicated by differences in slope T/C ratios. Continuous light stimulated fish growth during winter to spring, when the natural day length was short, without significantly changing white muscle and oocyte qualities in the fish of about 500 g, except for significantly increased white muscle RNA concentration. Continuous light also reduced fish growth rate later during summer, when the natural day length was long, by precedently decreasing the T/C ratio in late spring. Interestingly, plasma levels of free lysine related to tryptic digestion were correlated with trypsin specific activity levels. Continuous light caused higher levels of most free amino acids (FAA) involved in nitrogen metabolism, higher incorporation of essential FAA for protein synthesis, and higher protein turnover rate (free hydroxyproline levels) in both plasma and white muscle. However, continuous light did not affect higher protein content, intracellular buffering capacity and RNA levels in the white muscle of the fish of about 1 kg, probably due to limitation of FAA available for protein synthesis. It is therefore suggested that enhancing fish growth by continuous light stimulation should be accompanied by increasing availability or content of dietary protein (and probably minerals), which in turn would improve the quality of fish growth performance through increasing fillet protein concentration, strengthening vertebral growth, and delaying oocyte development.     

Effects of feeding frequency and dietary protein levels on juvenile allogynogenetic gibel carp (Carassius auratus gibelio) var. CAS III: growth,feed utilization and serum free essential amino acids dynamics

A 8 week growth trial of three feeding frequencies (2, 4 and 6 meals day^?1) and three dietary protein levels (30%, 34% and 38%) was conducted to investigate the influence of feeding frequency and dietary protein level on growth, feed utilization, serum free essential amino acids (EAA) dynamics and nitrogen (N), phosphorus (P), carbon (C) retention in juvenile allogynogenetic gibel carp (Carassius auratus gibelio) ‘CAS III’. The results showed that feeding frequency and dietary protein level increased feed intake (FI), specific growth rate (SGR) and feed efficiency (FE). SGR increased significantly with the increase in feeding frequency and dietary protein level. But, the increase in dietary protein level from 30% to 38% in group 6 meals day^?1 did not result in better growth. Compared with the group of 4 meals day^?1 × 38%, the fish fed the diet containing 30% crude protein at 6 meals day^?1 obtained similar SGR but showed lower FIp (protein intake). Compared with 2 meals day^?1, NREs (nitrogen retention efficiency) of 4 and 6 meals day^?1 were relatively higher. PRE (phosphorus retention efficiency) and CRE (carbon retention efficiency) increased significantly with feeding frequency while TNW (total nitrogen waste output), TPW (total phosphorus waste output) and TCW (total carbon waste output) decreased significantly. Feeding frequency and dietary protein level increased trypsin activity. Serum free EAA concentrations varied more synchronously at 6 meals day^?1 than 2 meals day^?1. It could be concluded that increasing feeding frequency could improve growth by increasing feed intake and feed efficiency for juvenile gibel carp. The increase in feeding frequency showed protein‐sparing effect.     

Chemical changes in fed and starved larval trout cod, Maccullochella macquarensis during early development

The changes in proximate composition, amino acid (total and free) and fatty acid content of artificially propagated trout cod, Maccullochella macquariensis larvae from five mothers hatched, weaned and reared separately, each in two groups, one fed with Artemia naupli and the other starved, for 15 days (after yolk resorption), are presented. There was no significant change in the proximate composition of fed larvae with devlopment, but in starved larvae the protein (linearly) and lipid (curvi-linearly) content decreased significantly as starvation progressed. The essential amino acids (EAA) and non- essential amino acids (NEAA) found in highest amounts in trout cod larvae were lysine, leucine, threonine and arginine, and alanine, serine and glutamic acid, respectively. In fed larvae the total amino acid (TAA), TEAA and TNEAA content did not vary significantly as development progressed. In starved larvae the TAA, EAA and NEAA content, as well as all the individual amino acids decreased significantly (P<0.05) from the levels in day of hatch and/or yolk-sac resorbed larvae. The greatest decrease occurred in the TEAA content (7.38±0.76 at day of hatch to 1.96±0.09 15 day starved in moles larva^–1; approximately a 74% decrease), whereas the decrease in TNEAA was about 38%. Unlike in the case of TAA distinct changes in the free amino acid (FAA) pool were discernible, from day of hatch and onwards, in both fed and starved trout cod larvae. In both groups of larvae the most noticeable being the decrease of % FEAA in TFAA, but not the % FAA in TAA. Four fatty acids together, accounted for more than 50% of the total in each of the major fatty acid categories in all larvae sampled; 16: 0, 18:1n-9, 22: 6n-3 and 20: 4n-6, amongst saturates, monoenes, n-3 PUFA and n-6 PUFA, respectively. Twelve fatty acids either decreased (14: 0, 16: 1n-7, 20: 1n-9, 20: 4n-6, 20: 5n-3, 22: 5n-3 and 22: 6n-3) or increased (18: 2n-6, 18: 3n-3, 18: 3n-6, 18: 4n-3 and 20: 3n-3) in quantity, after 15 days of feeding, from the base level in day of hatch and/ or yolk- sac resorbed larvae. The greatest increase occurred in 18: 3n-3 from 6.4±0.1 to 106.2±13.1 g mg lipid^–1 larva^–1, and the greatest decrease occurred in 22: 6n-3 (181.2±12.4 to 81.4±6.2 g mg lipid^–1 larva^–1). In starved larvae, at the end of 15 days, all the fatty acids, except 18: 0, 20: 3n-3 and 20: 4n-6, decreased significantly (P<0.05) from the levels in day of hatch and/or yolk- sac resorbed larvae.     

Effect of fish meal replacement with Carinata Brassica carinata in low animal protein diets of rainbow trout Oncorhynchus mykiss (Walbaum) on trypsin activity,protein and amino acid digestibility and bioavailability

Replacement of >25% of fish meal (FM) with aerobically converted carinata meal (ACCM) in low (200g/kg) animal protein (reference = 200g/kg FM) diets of rainbow trout Oncorhynchus mykiss resulted in reduced growth, partly due to reduced feed consumption and protein utilization. In this study, we determined the effect of FM replacement with ACCM on trypsin activity, apparent digestibility coefficients (ADCs) of protein, and essential amino acid (EAA) or conditionally essential amino acid (CEAA) ADCs and bioavailability. Replacement of FM did not alter protein ADCs or trypsin activity. Replacement of >25% FM reduced all EAA and CEAA ADCs except for arginine and leucine which were only reduced by 75% FM replacement. Only serum free lysine and muscle free histidine were reduced by >25% FM replacement. Muscle free lysine was only reduced by 75% FM replacement. Replacement of FM reduced EAA peak concentrations and resulted in slower release of EAAs in serum. Cumulative total EAAs in serum and muscle decreased with FM replacement. Ratios of EAAs to lysine showed that tryptophan was the most limiting EAA. However, isoleucine, leucine, methionine and phenylalanine were also inadequate for muscle synthesis for the first 9–12 hr following force‐feeding. Optimal time for protein synthesis was ≥36 hr. Although any level of FM replacement did not reduce protein ADCs and trypsin activity, replacement of ≥25% FM reduced EAA ADCs and bioavailability of lysine and histidine, which partly contributed to the observed differences in growth.     

Effects of dietary essential amino acid deficiencies on the growth performance and humoral immune response in silvery‐black porgy (Sparidentex hasta) juveniles

A 6‐week feeding trial was conducted for determining the effects of dietary essential amino acids (EAA) deficiencies on growth performance and non‐specific immune responses in silvery‐black porgy juveniles (4.7 ± 0.1 g initial weight). Eleven isoproteic (ca. 47%) diets were formulated including a control diet containing the optimum quantity of EAA, and ten EAA‐deficient diets. All diets contained 36% fish meal and 18.5% crystalline EAA and non‐essential amino acids (NEAA) as the main source of dietary proteins. All the EAA and NEAA incorporated in the crystalline amino acids mixture of the control diet simulated the amino acids profile of the fish meal. The other 10 EAA‐deficient diets were formulated by the deletion of each of the 10 EAA (crystalline form) from the control diet and replaced by a mixture of NEAA for the adjustment of dietary nitrogen contents. At the end of the experiment, fish fed with threonine‐deficient diet showed the lowest survival rate (p < .05), whereas growth performance decreased in fish fed all EAA‐deficient diets, although the reduction in body growth varied depending on the EAA considered. Plasma total protein decreased in all experimental groups except for fish fed the phenylalanine‐deficient diet. Fish fed with arginine‐ and lysine‐deficient diets had the lowest plasma C3, C4, lysozyme, total immunoglobulin and total superoxide dismutase activity (p < .05). Present results indicated that lysine, methionine and threonine were the most limiting EAA in terms of growth performance; however, arginine, threonine and lysine were the most limiting EAA for innate immunity responses in silvery‐black porgy juveniles.     

Evaluation of feed protein quality by measuring plasma free amino acids in Atlantic salmon (Salmo salar L.) after dorsal aorta cannulation

Two successive experiments were conducted in order to assess plasma free amino acid (FAA) profiles as a method for evaluating protein quality of fish feeds for Atlantic salmon, Salmo salar (L.). In experiment 1, the importance of meal size and inter‐fish variation was assessed by using dorsal aorta cannulated fish and diets that contained different sources of fishmeal (menhaden versus herring) which in the case of herring, had been dried at either 70 or 100 °C. In experiment 2, an attempt was made to mimic a production situation by comparing the FAA profiles in salmon fed diets containing two commercially available fishmeals that had been produced in accordance with industrial standards (Norse‐LT94® and NorSeaMink®; Norsildmel AL, Fyllingsdalen, Norway). FAA profiles in plasma 6 h after feeding were compared with feed true protein digestibility as determined in mink. Cannulated fish, held in individual tanks, were hand‐fed twice daily to pellet rejection (satiety) and daily records of the actual rations consumed were maintained. A total of 24 different amino acids and other amino‐containing compounds were detected using high‐pressure liquid chromatography. Morning and evening meal size showed significant correlations. Meal size had a significant effect on blood levels of the majority of essential free amino acids (EAA) as well as the total sum of FAA (TFAA). In experiment 1, a marked inter‐individual effect was found, possibly because of incipient sexual maturation. FAA profiles were therefore corrected for meal size by linear regression while repeated sampling via the permanently implanted cannula allowed paired comparisons of the different test diets, minimizing inter‐individual variation. Significant differences in plasma FAA profile, EAA and TFAA were detected between fish fed all diets in both experiments. The preceding parameters for fish ingesting each feed were directly related to their respective mink protein digestibility in experiment 1, but not in experiment 2. Our results show that dietary protein quality can be differentiated by the aforementioned protocol, and by using fish with a low metabolic rate feed qualities could be ranked correctly as in experiment 1.     

Effect of crude oil extracts from trout offal as a replacement for fish oil in the diets of the Australian native fish Murray cod Maccullochella peelii peelii

The efficacy of trout oil (TO), extracted from trout offal from the aquaculture industry, was evaluated in juvenile Murray cod Maccullochella peelii peelii (25.4±0.81 g) diets in an experiment conducted over 60 days at 23.7±0.8 °C. Five isonitrogenous (48% protein), isolipidic (16%) and isoenergetic (21.8 kJ g^?1) diets, in which the fish oil fraction was replaced in increments of 25% (0–100%), were used. The best growth and feed efficiency was observed in fish fed diets containing 50–75% TO. The relationship of specific growth rate (SGR), food conversion ratio (FCR) and protein efficiency ratio (PER) to the amount of TO in the diets was described in each case by second‐order polynomial equations (P<0.05), which were: SGR=–0.44TO²+0.52TO+1.23 (r²=0.90, P<0.05); FCR=0.53TO²–0.64TO+1.21 (r²=0.95, P<0.05); and PER=–0.73TO²+0.90TO+1.54 (r²=0.90, P<0.05). Significant differences in carcass and muscle proximate compositions were noted among the different dietary treatments. Less lipid was found in muscle than in carcass. The fatty acids found in highest amounts in Murray cod, irrespective of the dietary treatment, were palmitic acid (16:0), oleic acid (18:1n‐9), linoleic acid (18:2n‐6) and eicosapentaenoic acid (20:5n‐3). The fatty acid composition of the muscle reflected that of the diets. Both the n‐6 fatty acid content and the n‐3 to n‐6 ratio were significantly (P<0.05) related to growth parameters, the relationships being as follows. Percentage of n‐6 in diet (X) to SGR and FCR: SGR=–0.12X²+3.96X–32.51 (r²=0.96) and FCR=0.13X²–4.47X+39.39 (r²=0.98); and n‐3:n‐6 ratio (Z) to SGR, FCR, PER: SGR=–2.02Z²+5.01Z–1.74 (r²=0.88), FCR=2.31Z²–5.70Z+4.54 (r²=0.93) and PER=–3.12Z²–7.56Z+2.80 (r²=0.88) respectively. It is evident from this study that TO could be used effectively in Murray cod diets, and that an n‐3:n‐6 ratio of 1.2 results in the best growth performance in Murray cod.