Content of total and free amino acids in zooplanktonic food of fish larvae

The content of total and free amino acids in freshwater planktonic rotifers (Brachionus sp.), copepods (Eudiaptomus zachariasi) and groups of Daphnia pulex, Ceriodaphnia sp. and copepodites (Cyclops strenuus) of different sizes was determined. The amino acid content of Artemia salina nauplii on hatching and during fasting was also determined.

Amino acid content was lowest in rotifers and highest in stage III–IV copepodites. The major free amino acids in Cyclops strenuus dry matter were 1.43% arginine, 0.22% histidine, 0.20% alanine, 0.15% glutamic acid and 0.11% lysine. Free arginine content decreased in the daphnids as they increased in size. The content of all free amino acids in fasting Artemia nauplii was lower than in the freshwater zooplankters. The major free amino acids in nauplii were 0.55% proline, 0.41% alanine, 0.34% glycine and 0.37% serine. The content of most free amino acids in nauplii decreased during fasting. The significance of the results are discussed in relation to essential amino acid requirements of fish, and nutrition of fish larvae without fully developed gastrointestinal systems.     

Elution and internal migration of free amino acids in fish meats by soaking in sodium chloride or sorbitol solution

ABSTRACT:   Elution and internal migration of free amino acids (FAA) in fish meats by soaking were investigated when the meat strips were soaked in various concentrations of sodium chloride (NaCl) or sorbitol solution. Rapid decrease of FAA in an earlier phase of soaking was followed by a mild one, irrespective of the kind and the concentration of soaking solution. The loss of FAA by soaking in NaCl solution was slightly larger than that in sorbitol solution. However, regardless of the kinds of soaking solution, the elution of FAA from fish meats by soaking was only dependent on the soaking time, independently of the concentration, namely osmotic pressure of the soaking solution. Conversely, sluggish migration of FAA from the inside to the surface of the meat strips proceeded by soaking. These results suggested that the elution of FAA from fish meats by soaking was driven by a simple diffusion from the surface of the meats to soaking solution, but it was regulated by sluggish migration rate of FAA in fish meats.     

Culture-independent analysis of the bacterial community during fermentation of pa-som,a traditional fermented fish product in Laos

Pa-som is a traditional salty fermented food made from varieties of freshwater fish with garlic and rice in Laos. A similar product, plaa-som, is made in Thailand. To investigate the fermentation process of pa-som, lactic acid production and pH were monitored during laboratory-scale production of pa-som. Furthermore, the variety of bacteria involved and their succession during fermentation were examined using culture-independent polymerase chain reaction (PCR) denaturing gel gradient electrophoresis targeting the 16S ribosomal RNA (rRNA) gene. d- and l-lactic acid production and a decrease in pH were observed in the samples after 1 day of fermentation. The lactic acid content and pH continued to increase and decrease, respectively, until 4 days of fermentation. Overall, six lactic acid bacteria and eight other indigenous bacteria species were detected during pa-som fermentation. Among the lactic acid bacteria detected, two Lactococcus and one Weissella species were detected in samples after 1–4 days of fermentation, suggesting that these three species play major roles in pa-som fermentation from the initial phase. This is the first report to investigate the fermentation process in pa-som at the microbial level. This research approach should be applicable to a wide variety of pa-som and related fermented fishery products in Laos to help understand their microbial diversity and to identify beneficial bacterial species for improving quality.     

杂交鳜与鳜鱼、斑鳜肌肉营养成分和氨基酸含量比较

通过测定鳜鱼、斑鳜和杂交鳜的肌肉营养成分和18种氨基酸含量,结果表明：杂交鳜、斑鳜和鳜鱼的肌肉水分、脂肪、灰分的含量无显著差异（P〉0．05）,杂交鳜与斑鳜的肌肉蛋白质含量无显著差异（P〉0．05）,但均显著高于鳜鱼肌肉的蛋白质含量（P〈0．05）;鳜鱼、斑鳜和杂交鳜的18种氨基酸含量及其组成、18种氨基酸总量（TAA）、人体所需8种必须氨基酸总量（HEAA）及4种呈味氨基酸（天门冬氨孽、谷氨酸、甘氨酸和丙氨酸）的总量（FAA）均无显著差异（（P〉0．05）。     

杂交罗非鱼饲料中豆粕、发酵豆粕和晶体氨基酸替代鱼粉的研究

添加豆粕(soybean meal,SBM)、发酵豆粕(fermented soybean meal,FSBM)和晶体氨基酸(crystalline amino acid,CAA)替代杂交罗非鱼(Oreoehromis niloticus ♀×O.aureus ♂)实用饲料中5%的鱼粉,连续投喂初始体重为9.41±0.05 g的罗非鱼9周后,通过测定罗非鱼的增重率(WGR,%)、特定生长率(SGR,%)、饲料效率(FE,%)、蛋白质效率(PER,%),并进行肝脏分析和血浆分析,比较各组饲料对罗非鱼生长和非特异性免疫的影响.结果表明,发酵豆粕+晶体氨基酸组的WGR、SGR高于其他3组,鱼粉组的FE高于其他3组;PER方面各组间差异不显著,4组饲料对罗非鱼肝脏组成、血浆成分和存活率没有造成显著差异.     

细角螺肌肉氨基酸及脂肪酸组成分析

测定了细角螺(Hemifusus termatamus)腹足部肌肉的氨基酸含量与脂肪酸组成,并与方斑东风螺(Babylonia areolata)、管角螺(Hemifusus tuba)、棘角螺(Hemifusus kawamurai)和鲐鱼(Pneumatophorus japonicus)进行了比较。结果显示,细角螺肌肉中检出17种氨基酸,总含量为肌肉干重的81.03%,高于鲐鱼(79.86%)、方斑东风螺(66.45%)、管角螺(56.94%)和棘角螺(64.42%);肌肉中4种呈味氨基酸(天冬氨酸、谷氨酸、甘氨酸和丙氨酸)的含量为38.49%,占氨基酸总量的47.50%;8种必需氨基酸(EAA)总含量为38.49%,占氨基酸总量的46.94%。细角螺肌肉中共检出20种脂肪酸,其中饱和脂肪酸的含量为65.25%,不饱和脂肪酸的含量为34.75%,不饱和脂肪酸与饱和脂肪酸含量的比值为0.53,低于方斑东风螺(1.15)、管角螺(1.13)和棘角螺(0.69),高于鲐鱼(0.37)。结果表明,细角螺肌肉中必需氨基酸与不饱和脂肪酸含量较丰富,具有较高的营养价值,是一种较为理想的食物蛋白与脂肪来源。     

锈斑蟳肌肉氨基酸和脂肪酸组成分析及营养品质评价

测定了锈斑蟳(Charybdis feriatus)肌肉的氨基酸含量与脂肪酸组成,对其氨基酸含量和脂肪酸组成进行了分析和评价。结果显示,锈斑蟳肌肉干样中共检测出包括牛磺酸在内的19种氨基酸,组成蛋白质的氨基酸总含量为肌肉干重的75.36%,其中,8种必需氨基酸(EAA)的总量为25.41%,占氨基酸(AA)总量的33.73%,非必需氨基酸(NEAA)总量为40.91%,而必需氨基酸与非必需氨基酸的比值(EAA/NEAA)为62.13%;肌肉中4种鲜味氨基酸的含量为28.84%,占氨基酸总量的38.28%;锈斑蟳肌肉中检测出了12种脂肪酸,其中,饱和脂肪酸(SFA)的含量为38.04%,单不饱和脂肪酸(MUFA)的含量为31.58%,多不饱和脂肪酸(PUFA)的含量为29.80%,多不饱和脂肪酸与饱和脂肪酸比值(PUFA/SFA)为0.78,n-3 PUFA与n-6PUFA的比值达到了17.75,多不饱和脂肪酸中EPA和DHA两者的含量达到了20.24%。结果表明,锈斑蟳肌肉中必需氨基酸、鲜味氨基酸含量与不饱和脂肪酸比例较高,具有较高的营养保健价值。     

Changes in the levels of circulating plasma free amino acids of carp (Cyprinus carpio) after feeding a protein and an amino acid diet of similar composition

A protein diet containing casein as the sole nitrogen source and an amino acid diet of similar composition (except isolated amino acids represented all of the nitrogen) were fed to triplicate groups of yearling carp. The sequence of appearance of free amino acids in the plasma of carp at 0, 2, 4, 8 and 16 h after feeding the two diets was examined.The circulating levels of all plasma free essential amino acids were in highest concentrations at 4 h after feeding the casein diet, except for arginine which declined slightly from 2 to 4 h. After feeding the amino acid diet, plasma free arginine and lysine were at peak levels at 2 h after feeding, while others peaked by 4 h.Glutamic acid and aspartic acid were detected in very low concentrations in the plasma relative to their levels in the diets, while proline was found in the highest concentrations of all other amino acids.Essential amino acid levels in the plasma were found to be positively correlated with their levels in the diets, while non-essential amino acids were not.     

Hydrolyzates produced from mackerel Scomber japonicus skin by the pressurized hydrothermal process contain amino acids with antioxidant activities and functionalities

The production of amino acids with antioxidant activities and functional properties from protein hydrolyzates of freeze-dried and supercritical carbon dioxide (SC-CO₂)-extracted mackerel skin by pressurized hydrothermal hydration (PHH) at different temperatures (150–240 °C) and pressures (12–210 bars) was investigated. The highest yield of amino acid in freeze-dried and SC-CO₂-extracted mackerel skin hydrolyzates was 121.93 ± 1.80 and 122.96 ± 2.84 mg/g, respectively, at 240 °C and 210 bars. Nine essential amino acids were identified in both skin hydrolyzates, of which histidine was the most abundant. All essential amino acids showed a temperature stability up to 240 °C, with the exception of threonine and histidine. The antioxidant activity of the hydrolyzates, as demonstrated in the DPPH, ABTS, hydroxyl radical, metal chelating, and reducing power assays, increased with increases in temperature and pressure; it was high in both hydrolyzates at 240 °C and 210 bars. In terms of functional properties, hydrolysis at different temperatures and pressures increased protein solubility to above 59 % over a wide pH range (3.5–9.5). When the temperature and pressure increased, the emulsifying activity index, emulsion stability, foaming capacity, and foam stability of both hydrolyzates decreased, possibly caused by the shorter peptide chain length. We conclude that protein hydrolyzates produced from mackerel skin can be used in food-related industries as good additives.     

Isolation and characterization of a rhabdovirus from co-infection of two viruses in mandarin fish

Co-infection of two viruses has been observed in mandarin fish (Siniperca chuatsi), but the two viruses have not been characterized. In this study, a rhabdovirus has been isolated from the co-infected two viruses extracted from the diseased mandarin fish, and its morphological structure and partial biochemical and biophysical characteristics have been observed and analyzed. The isolated rhabdovirus has a typical bullet shape, and is therefore called S. chuatsi rhabdovirus (SCRV). And, the isolated rhabdovirus produced a higher titer (10^8.5 TCID₅₀ ml^− 1) than did the co-infecting viruses (10^6.5 TCID₅₀ ml^− 1). Subsequently, the viral genome RNA was extracted, and used as template to clone the complete nucleoprotein (N) gene by RT-PCR amplification. Cloning and sequencing of the SCRV N protein revealed 42%-31% amino acid identities to that of trout rhabdovirus 903/87 and the rhabdoviruses in genus Vesiculovirus. SDS-PAGE separation of the isolated SCRV and other two rhabdoviruses also revealed obvious polypeptide profile difference. Moreover, the anti-SCRV N protein antibody was prepared, and the anti-SCRV N protein antibody only could recognize the SCRV N protein, whereas no antigenicity was detected in other two rhabdoviruses. The data suggested that the SCRV should be a rhabdovirus member related to the genus Vesiculovirus in the Rhabdoviridae.     

Absorption, assimilation and catabolism of individual free amino acids by larval Atlantic halibut (Hippoglossus hippoglossus)

Following the completion of the lecithotrophic phase, most marine larvae rely on an incompletely developed digestive tract to absorb amino acids (AA) and other nutrients needed for rapid growth. Despite their undeveloped state, larvae must absorb nutrients in sufficient amounts to fuel exceptionally high rates of growth. This study examined the ability of larval Atlantic halibut to absorb, assimilate and catabolise dispensable (alanine, glutamate) and indispensable (arginine, lysine) dietary free amino acids (FAA) using tube-fed AA solutions with ¹⁴C tracers. Absorption of FAA was rapid with an average of 71% absorbed from the gut within 30 min after tube feeding. Evacuation of FAA by larvae was low, averaging only 6% of tube-fed dose. Dispensable amino acids (DAA) were catabolised in greater proportion (17%) than indispensable FAA (54%). Saturation of FAA transporters was not attained under the present conditions (20 mM, 20% midgut filling). Absorption rates did not differ significantly between the four FAA.     

Pilot study of the relationship of free amino acids in serum and in the cerebrospinal fluid of horses

In a blind study serum and cerebrospinal fluid (CSF) of control horses and of horses in hepatic coma after chronic food intoxication with Senecio alpinus were collected simultaneously and the composition of free amino acids was determined. The hepatic encephalopathy index in serum (less than 1.65) and in CSF (less than 1.11) of liver patients was distinctly less than to the control values in serum (greater than 2.42) and in CSF (greater than 1.49). The serum concentrations of glutamic acid in hepatic coma were elevated five-fold in comparison to the controls. An indication of ammonia decontamination was that nearly ten-fold higher values of glutamine were found in the cerebrospinal fluids of patients than in serum. In comparison to controls the serum levels of glutamine in horses with hepatic encephalopathy were decreased by the factor 0.7.     

Seasonal variations of free amino acids and nucleotide-related compounds in the muscle of cultured Taiwanese puffer Takifugu rubripes

SUMMARY: Seasonal variation in the level of extractive nitrogenous components was investigated in the muscle of puffer Takifugu rubripes cultured in two different areas of Taiwan. There were no seasonal and regional variations in the proximate composition of the fish. Of the free amino acids (FAA) in the muscle of puffer, the predominant one was taurine, followed by glycine, lysine, and alanine. Among nucleotide-related compounds, inosine monophosphate (IMP) and guanosine monophosphate (GMP) were the most prominent compounds. The total nucleotide-related compounds in the muscle of puffer were higher in July and November than those in the other months. The level of total taste-active components including glycine, alanine, arginine, GMP, IMP, and adenosine 5'-monophosphate was much higher in the muscle of puffer collected from July to January. Therefore, the puffer is probably more palatable in these periods.     

Periodical changes of plasma free amino acid levels and feed digesta in yellowtail after feeding non-fishmeal diets with or without supplemental crystalline amino acids

ABSTRACT: Postprandial changes of free amino acid (FAA) concentrations in plasma and feed digesta contents were examined in yellowtail, Seriola quinqueradiata (220–280 g bodyweight) fed non-fishmeal diets either with or without supplemental crystalline amino acids (lysine, methionine, threonine, and tryptophan) in order to evaluate the availability of supplemental amino acids. Non-fishmeal diets containing 30% soy protein concentrate as the major protein source were prepared in three diet forms: soft dry pellet, extruded pellet, and single moist pellet. The level of plasma FAA and feed digesta content were determined at 0, 3, 6, 9, 12, 18, and 24 h after feeding. Plasma levels of four supplemental amino acids in fish fed the non-fishmeal diets with EAA were higher than those of fish fed diets without EAA, suggesting that yellowtail can efficiently absorb supplemental crystalline amino acids irrespective of diet form. However, a remarkable difference was observed in the periodical patterns of these four amino acids between FAA derived from supplements and those from feed protein. Moreover, FAA patterns in fish fed the non-fishmeal diets with EAA were different from those of the control fishmeal diet. Of note, methionine concentration was markedly high during the whole experimental period, resulting in an amino acid imbalance that may have caused lower feed performances in fish fed the non-fishmeal diets with EAA compared to the control fishmeal diet.     

银鲳与翎鲳肌肉必需氨基酸组成模式的比较分析

比较分析了相同规格银鲳与翎鲳幼鱼肌肉必需氨基酸的组成模式。结果显示,银鲳与翎鲳肌肉中氨基酸均以谷氨酸含量最高,分别为2.91、2.77 g/100 g湿物质,色氨酸含量最低,分别为0.17、0.18 g/100 g湿物质;银鲳肌肉中氨基酸总量、必需氨基酸总量及呈味氨基酸总量分别为18.08、9.32与6.84 g/100 g湿物质,翎鲳肌肉中氨基酸总量、必需氨基酸总量及呈味氨基酸总量分别为17.25、8.80与6.60 g/100 g湿物质;银鲳肌肉中必需氨基酸的组成比例为缬氨酸∶蛋氨酸∶异亮氨酸∶亮氨酸∶苏氨酸∶苯丙氨酸∶组氨酸∶赖氨酸∶精氨酸∶色氨酸=6.02∶3.12∶5.66∶9.45∶4.33∶4.50∶2.56∶11.15∶7.06∶1.00,翎鲳肌肉必需氨基酸中缬氨酸∶蛋氨酸∶异亮氨酸∶亮氨酸∶苏氨酸∶苯丙氨酸∶组氨酸∶赖氨酸∶精氨酸∶色氨酸=5.64∶2.42∶5.18∶8.74∶4.12∶4.13∶2.46∶10.28∶6.65∶1.00。统计分析表明,银鲳与翎鲳间肌肉中氨基酸总量、必需氨基酸总量、呈味氨基酸总量以及必需氨基酸的组成模式并无显著性差异(P0.05)。     

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.     

Utilization of free amino acids, yolk proteins and lipids in developing eggs and yolk-sac larvae of walleye pollock Theragra chalcogramma

ABSTRACT:   To elucidate the utilization of the major yolk nutrient stocks in eggs and larvae of walleye pollock Theragra chalcogramma , the contents of free amino acids (FAA), the major yolk protein (180 kDa lipovitellin originated from vitellogenin B in ovulated eggs: oLv B), and lipids were measured. Most eggs hatched 18 days after fertilization at 5°C, and all larvae absorbed almost all their yolk mass by 28 days. The total FAA content showed no change during the first 6 days, and then decreased to 28% of the initial level by 18 days. The oLv B contents, measured by an enzyme-linked immunosorbent assay using a specific antiserum against oLv B, gradually decreased from 6 to 18 days, followed by a rapid decline. The content of phospholipids (PL) and triacylglycerols (TG) showed no marked change until hatching, and then decreased until disappearance of yolk sac. From these results, it is proposed that there are two main periods for nutrient utilization in embryos and larvae of walleye pollock. In the first period, FAA was mainly utilized until 18 days after fertilization. Active utilization of oLv B and lipids (PL and TG) instead of FAA occurred during the second period from 18 to 28 days.     

The patterns of plasma free amino acids after force-feeding in rainbow trout Oncorhynchus mykiss (Walbaum) with and without dorsal aorta cannulation

Two experiments were conducted to compare the patterns of plasma free amino acid concentrations after force-feeding in rainbow trout Oncorhynchus mykiss (Walbaum) with and without dorsal aorta cannulation. In the first experiment, 35 rainbow trout averaging 504 ± 7.8 g (mean ± SD) were divided into seven groups of five fish each. After 48 h starvation, a group of fish was anaesthetized and blood samples were taken at one of the following time periods: 0, 4, 8, 12, 24, 36 and 48 h after feeding. In the second experiment, five dorsal aorta cannulated rainbow trout averaging 511 ± 6.2 g (mean ± SD) were kept in a cage. After 48 h starvation, the fish were anaesthetized and blood samples were taken from the same fish at 0, 4, 8, 12, 24, 36 and 48 h after feeding. In the first experiment, the concentration of all plasma free amino acids except histidine and glycine peaked at 4 h and returned to the basal level 24 h after feeding. In the second experiment, the concentration of all plasma free amino acids except isoleucine, leucine, phenylalanine and tryptophan also peaked at 4 h and returned to the basal level 24 h after feeding. These results showed that the pattern of plasma free amino acid concentrations from fish with and without dorsal aorta cannulation were similar.     

A comparison of two faecal collection methods for protein and amino acid digestibility coefficients of menhaden fish meal and two grades of poultry by-product meals for market-size sunshine bass (Morone chrysops × M. saxatilis)

Apparent digestibility and availability coefficients for protein and amino acids in menhaden fish meal (MEN), pet‐food grade (PBM‐pet,) and feed‐grade poultry by‐product meal (PBM‐feed) were determined for market‐size (500 g) sunshine bass in two consecutive trials using passive netting (1.6 mm mesh) followed by manual stripping of faeces. A reference diet resembling a commercial feed was formulated to meet or exceed all known nutritional requirements of hybrid striped bass. Test diets were formulated to contain a 70 : 30 mixture of reference diet to test ingredient with chromic oxide (1%) serving as the inert marker. Diets were extruded under commercial conditions and the reference diet was fed for two weeks in order to acclimate fish to experimental conditions. Each diet was randomly assigned to triplicate tanks containing 30 fish each. Fish were fed their respective diet twice daily to apparent satiation for two weeks, with faecal collections being conducted on the 7th and 14th day, in each trial. ADC values determined in the net method were highly variable and generally lower than ADCs obtained by stripping. Consistently lower and highly variable chromium concentrations were found in the net method faecal samples and suggest that marker loss relative to nutrient content, or dilution of marker with non‐faecal matter, on the net collectors influenced results in that trial. In contrast, the standard errors of ADCs determined in the strip method were less than 5 percentage points in most cases. Protein digestibility ranged from a low of 51% (PBM‐feed) to a high of 87% (PBM‐pet) in the net method, and from a low of 80% (PBM‐feed) to a high of 99% (MEN) in the strip method. With the exception of Lys, no differences in amino acid availabilities among diets were found in the net method. In the strip method, protein digestibility and amino acid availabilities in MEN were generally greater than those found in PBM‐pet or PBM‐feed, whereas ADCs were not significantly different between the two poultry by‐products. Based on the conditions of the present study, net collection of faecal matter can not be recommended for determining the digestibility of nutrients in feed ingredients for market‐size sunshine bass. Digestibility coefficients obtained by the strip method for feed and petfood grades poultry by‐product were higher than those previously reported and will be facilitate more efficient and economical diet formulations for larger sunshine bass.