C-protein (MyBP-C) isoforms from carp ordinary and dark muscles and muscle type-specific binding to myosin

ABSTRACT:   C-protein is a myosin-associated protein of vertebrate striated muscle, and its function and properties have been extensively examined. However, there has been no report of C-protein of fish skeletal muscle so far. C-protein was identified in carp skeletal muscle by immunoassay using antibody against chicken C-protein, and the muscle-type specific C-protein was purified from carp ordinary and dark muscles for the first time. Although C-protein could be prepared from crude myosin by the reported procedure, C-protein degraded appreciably during the purification steps. Accordingly, C-protein was selectively extracted from the muscle with 0.15 M K-phosphate buffer (pH 5.8), and purified by ammonium sulfate fractionation, followed by AF-blue chromatography. Myosin free from the accessory proteins was obtained by diethylaminoethyl (DEAE) chromatography and used to assay the binding of C-protein with myosin. Ordinary muscle C-protein bound to ordinary muscle myosin in a saturable manner, but its maximum amount of binding was approximately twice that of dark muscle myosin. Similarly, dark muscle C-protein bound to dark muscle myosin much more than to ordinary muscle myosin. These results suggest that C-protein isoforms specifically bound with myosin isoforms originated from the same type of muscle.     

Influence of interposition of pink muscle fiber into dorsal ordinary muscle on 5'-IMP degrading activity in various fish species

ABSTRACT: To clarify the cause of differences in the temporal change of K -value among fish species living in the same habitat water temperature, the influence of the interposition of pink muscle fibers into dorsal ordinary muscle on 5'-inosine monophosphate (5'-IMP) degrading activity was examined. Fourteen fish species from the Sakishima Islands (habitat water temperature 28°C) and from Nagasaki (habitat water temperature 17°C) were used for the sample fishes. Each of 5'-IMP and ρ-nitrophenol phosphate (ρ-NPP) degrading activities showed a peak at near pH 8.0 and near pH 5.0, respectively. These activities were somewhat higher in fish from Nagasaki than in fish from the Sakishima Islands. The interposition percentage of pink muscle fibers into dorsal ordinary muscle correlated significantly ( P < 0.05) with the 5'-IMP degrading activity at pH 7.0. Furthermore, the activity at pH 7.0 correlated significantly ( P < 0.001) with the increasing rate of K -value at 32°C (ΔK₃₂). These results suggest that the difference in ΔK₃₂ among fish species in the same habitat water temperature might be caused by differences in 5'-IMP degrading activity because of the differences in the interposition rates of pink muscle fibers into dorsal ordinary muscle.     

Preventive method of color deterioration of yellowtail dark muscle during frozen storage and post thawing

ABSTRACT:   The aim of this study was to develop a practical preventive method for color deterioration of sliced or filleted yellowtail muscle, especially of dark muscle, during frozen storage and post thawing. When the sliced meats were packaged in a vacuum with low oxygen permeable flexible films and then stored frozen below −40°C, no significant discoloration or browning of dark muscle was observed for 9 months or more. For higher temperature storage at −20°C or −30°C, nitrogen gas substituted packaging was a useful practical method for storing sliced meats for 6 weeks. In order to prevent color deterioration of sliced meats after thawing and subsequent storage at 0°C, the efficacy of materials of packaging was investigated. The most desirable result was obtained by using a film with oxygen permeability of 50–90 cm³/m² per 24 h.     

Replacement of dietary fish oil with olive oil in young yellowtail <Emphasis Type="Italic">Seriola quingqueradiata</Emphasis>: effects on growth,muscular fatty acid composition and prevention of dark muscle discoloration during refrigerated storage

Dietary fish oil (FO) was replaced by olive oil (OO) in young yellowtail Seriola quinqueradiata to investigate its effects on growth, muscular fatty acid composition and prevention of color deterioration of dark muscle during storage. Yellowtail were fed one of four diets, where FO (80 g/kg diet) was replaced by OO (0, 25, 50 and 100%) for 40 days. No significant difference in growth was seen among the diet groups. In addition, these experimental diets did not affect the proximate compositions of the dorsal muscle, ventral muscle and the liver in these fish. Serum total protein, glucose and total cholesterol levels did not show significant differences; however, serum triglyceride levels were significantly higher in fish fed 50 and 100% OO diets. Fatty acid composition of ventral muscle reflected the composition of the respective diets. Dark muscle discoloration was reduced in fish fed OO diets after 12 to 18 h during storage at 4°C. Furthermore, the redness value of fresh dark muscle increased depending on the extent of FO replacement. These results indicate that the partial or total dietary replacement of FO with OO prevents discoloration of dark muscle without affecting the growth of young yellowtail after 40 days of feeding.     

Influence of pH and temperature on force development and shortening velocity in skinned muscle fibres from fish

Three species of fish were studied: Atlantic cod (Gadus morhua), sculpin (Myoxocephalus scorpius) (from the North Sea, temperature 2 to 12°C) andNotothenia neglecta (from Antarctica, temperature –2 to +2°C). Single fast muscle fibres were isolated from anterior myotomes and skinned with detergent in order to directly determine the effects of pH and temperature on force production and shortening velocity.In all species maximum force production (Po) was independent of pH over the range 7.3–8.0. Decreasing the pH from 7.3 to 6.6 reduced maximum force by 28% in fibres fromG. morhua andN. neglecta but had no effect on fibres fromM. scorpius. The depression in maximum force with acidosis was accompanied by a proportional decrease in stiffness and an increase in the rate of force recovery after stretch.Unloaded contraction velocity of cod fibres (Vmax) showed a pH optimum at around pH 7.6 decreasing by 31% at pH 6.6. Vmax of fibres from the other species was independent of pH over the range 6.6–8.0.The effects of pH on Po and Vmax were similar at 0 and 10°C. Thus for maximally activated fibres both force and contraction velocity are independent of temperature induced changes in pH. In some species acidosis depresses contractility and is likely to be a contributory factor to muscle fatigue.     

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）。     

鱼贝类肌肉品质变化与能量代谢关联

水产养殖是满足人们对蛋白质需求的重要途径,减少鱼贝类捕后因鲜度下降导致的资源浪费是水产品加工领域的重要研究课题之一。横纹肌作为鱼贝类肌肉中主要的食用部位,其品质变化与肌肉能量代谢状况密不可分。本文详细综述了ATP在线粒体、肌质网和肌原纤维蛋白间的产生、转运和消耗的过程。回顾了基于横纹肌能量代谢过程所提出的一系列鱼贝类肌肉鲜度评价指标,包括ATP及其关联化合物相关的K值、K_max值、AEC值,以及肌原纤维蛋白和肌质网的ATPase活性、线粒体结构和呼吸活性等。进一步根据消费现状综述了各鲜度指标在鱼类鲜品和贝类活品中的适用性,提出加强鱼贝类捕后初期（品质易逝期）控制的必要性和紧迫性。在众多指标中,线粒体活性可作为易逝期品质控制的有效指标,可有效开展易逝期的品质评价,减少因捕后处置不当造成的品质损失和资源浪费。     

Changes in Volatile Compounds of Dark and Ordinary Muscles of Yellowtail (Seriola quinqueradiata) During Short-Term Cold Storage

ABSTRACT

Changes in volatile compounds in dark and ordinary muscle of yellowtail during 2 days of refrigerated storage at 5°C were investigated. Twenty-seven compounds were identified in ordinary muscle and 39 in dark muscle during 2 days of storage at 5°C. Thirteen compounds at Day 0 of storage and 29 compounds at Day 2—such as 2,3-pentadione, 1-penten-3-ol, (Z)-2-penten-1-ol, and (E,Z)-2,4-heptadienal—identified in the dark muscle showed significantly higher values compared to ordinary muscle. Levels of thiobarbituric acid-reactive substances (TBARS) in dark muscle were significantly higher than those in ordinary muscle throughout 2 days of storage period, and a significant increase in TBARS occurred in just dark muscle at Day 2 of storage. In ordinary muscle, viable cells remained at the same order of magnitude as their initial values for 2 days. Eight aldehydes in ordinary muscle and 25 volatile compounds in dark muscle increased significantly without microbial action prior to increase in TBARS during short-term cold storage. Principal component analysis of the volatile compounds in dark and ordinary muscle was able to differentiate between different storage time samples of same muscle type as well as different muscle samples of same storage time.     

工业化海水鱼类养殖现状与发展趋势

工业化养鱼是农业现代化发展的必然趋势,合乎国情的封闭式循环水养鱼是我国水产养殖的发展方向,配置科学合理的装备技术,通过水体颗粒物分离、控制病原菌数量、微生物净化、增氧等技术措施,使养殖环境达到生态清洁型,健康养殖,确保食品安全。     

三种不同游泳习性鱼类骨骼肌的快、慢肌纤维组织学特性

硬骨鱼类的骨骼肌可为其游泳运动提供动力。为认识不同游泳习性鱼类的骨骼肌肌纤维组织学特征,本研究选取3种具有不同游泳习性的鱼类：鲐(Scomber japonicus)、大黄鱼(Larimichthys crocea)和褐牙鲆(Paralichthys olivaceus),采用石蜡切片苏木精–伊红染色法和形态计量法,以形状、直径和密度作为评价指标,对其骨骼肌的快、慢肌纤维组织学特性进行表征,比较3种不同游泳习性鱼类的快、慢肌纤维组织学特征差异。组织学观察发现,3种类型鱼类的骨骼肌快、慢肌纤维横切面均呈不规则形状。营持续式游泳的鲐的快肌纤维呈多角状,慢肌纤维呈多边柱形;营延长式游泳的大黄鱼的快、慢肌纤维呈长椭圆形;营爆发式游泳的褐牙鲆的快、慢肌纤维呈扁椭圆形。形态计量结果显示,3种鱼类的快肌纤维直径均极显著大于其慢肌纤维(P<0.01)。大黄鱼的慢肌纤维直径在3种鱼类中最大,约为鲐的1.34倍、褐牙鲆的1.14倍;鲐的快肌纤维直径在3种鱼类中最大,约为大黄鱼的1.41倍、褐牙鲆的1.35倍。3种鱼类的快肌纤维密度均极显著小于其慢肌纤维(P<0.01),其中,快肌纤维密度大小排...     

Plasticity of the properties of mitochondria from rainbow trout red muscle with seasonal acclimatization

Cold-acclimation of rainbow trout brings only limited changes in muscle metabolic capacities, but marked modifications in membrane composition. Thus, we examined whether the functional properties of mitochondria from trout red muscle were modified by seasonal temperature acclimatization. Mitochondria from fall-acclimatized trout had higher maximal capacities (state 3 rates) for the oxidation of pyruvate and acyl carnitines at 12 and 20 °C than mitochondria isolated from summer-acclimatized trout. For these substrates, the increased oxidative capacity completely compensated for the seasonal drop in temperature. Pyruvate and palmitoyl carnitine were consistently the preferred substrates, while decanoyl and octanoyl carnitine were oxidized at higher rates than glutamine, particularly in fall trout. State 4 rates of oxygen uptake (obtained when all ADP has been converted to ATP) differed less among substrates, but varied seasonally. State 4 rates at 12 and 20 °C were higher in mitochondria isolated from fall than summer trout. At low temperatures, the Q₁₀ of both maximal and state 4 rates of substrate oxidation tended to be higher for mitochondria from fall trout. The apparent Arrhenius activation energy (E_a) for mitochondrial pyruvate oxidation was higher in fall than summer trout whereas the E_as for palmitoyl carnitine and decanoyl carnitine oxidation did not change. The fatty acids of mitochondrial phospholipids from fall trout were more polyunsaturated than those from summer trout, with 12% more double bonds occurring than in summer trout, suggesting that membrane restructuring may be involved in the observed compensatory responses.     

Anabolism,catabolism and proteomic analysis in the slow‐twitch muscle of pacu (Piaractus mesopotamicus) submitted to prolonged fasting followed by refeeding

Here, we aimed to study the slow muscle of the fish Piaractus mesopotamicus submitted to 30 days of fasting (D30) followed by 1 day (D31) or 30 days of refeeding (D60). Histological analysis of fibre diameter was performed in D30 and D60. The gene expression of parvalbumin (pvalb), atrogenes (murf1a, murf1b, mafbx) and anabolic genes (igf‐1, mtor) was analysed using RT‐qPCR in D30, D31 and D60. The proteome was obtained by shotgun proteomics at D30 and D60, and the set of differentially expressed proteins was analysed by bioinformatics. In all experiments, the control was regularly fed fish. The histological analysis showed no changes in muscle fibre diameter. The expression of catabolic and anabolic genes was not changed, except for the downregulation of igf‐1 in D30 and of mafbx in D31. The expression of pvalb was not changed in D30 and D60 but was decreased in D31. The proteomic analysis identified 169 proteins in D30 (24 upregulated and 18 downregulated) and 170 proteins in D60 (17 upregulated and 21 downregulated); many of them were related to energetic metabolism and intracellular Ca²⁺ homoeostasis. Overall, our results indicate that the slow‐twitch muscle presented few changes upon prolonged fasting and refeeding condition.     

Characterization of virulence,cell surface characteristics and biofilm‐forming ability of Aeromonas spp. isolates from fish and sea water

Members of the genus Aeromonas are emerging human pathogens, causing a variety of extra‐intestinal, systemic and gastrointestinal infections in both immunocompetent and immunocompromised persons. Aeromonas virulence is multifaceted and involves surface‐associated molecules, motility, biologically active extracellular products and biofilm formation. Aeromonads, isolated from diverse freshwater fish species as well as sea water, were screened for biofilm formation, with varying physicochemical parameters including temperature, agitation and nutrient availability. Motility, cell surface characteristics (auto‐aggregation, hydrophobicity and S layer), and extracellular virulence factor production (haemolysis, proteolysis, DNase production) were also assessed to identify potential associations with the biofilm phenotype. Biofilm formation was influenced by environmental conditions, with isolates preferentially forming biofilms in nutrient‐rich media at 30 °C, although strong biofilm formation also occurred at 37 °C. Strong biofilm formation was observed for Aeromonas culicicola isolates following exposure to nutrient‐rich conditions, while Aeromonas allosaccharophila isolates preferred nutrient‐poor conditions for biofilm formation. Source‐/species‐specific correlations, ranging from weak to strong, were observed between biofilm formation and motility, cell surface characteristics and/or extracellular virulence factor production. Understanding the specific mechanisms by which Aeromonas species adhere to abiotic surfaces may aid in preventing and/or treating disease outbreaks in aquaculture systems and could lead to effective eradication of these fish pathogens.     

Pyridinoline concentrations in muscular and skin collagen of fish and relationship between collagen solubility and pyridinoline concentration in fish muscular collagen

Pyridinoline (Pyr), one of the mature crosslinks of collagen, was determined in muscular collagen of three species of fish. The amounts of muscular Pyr in red sea bream, yellowtail, and tiger puffer were 3.4, 8.8, and 50.3 mmol/mol collagen, respectively, indicating that the Pyr concentration in muscular collagen differs greatly among fish species. The Pyr concentration in tiger puffer muscular collagen was the greatest, but it was only one-fourth that in rabbit muscle. As in mammalian skin collagen, Pyr was not detected in skin collagen of red sea bream and yellowtail. However, tiger puffer skin contained Pyr (3.75 mmol/mol collagen). The presence of Pyr would have a relationship to some features of tiger puffer skin, such as mechanical strength and thickness. Pyr concentrations in acid-soluble collagen (ASC), pepsin-solubilized collagen (PSC), and insoluble collagen (ISC) in muscles of three species of fish were determined. Pyr was found in ISC > PSC > ASC, from the highest to the lowest concentration, and the concentration in ISC was 45–200 times that in ASC. Therefore, Pyr crosslinks that are formed between collagen molecules would have a close relationship to collagen solubility.     

cDNA cloning and expression analysis of the myosin heavy chain (MYH) gene of the mandarin fish Siniperca kneri

In this study, we applied RT-PCR and cDNA cloning techniques to clone myosin heavy chain (MYH) cDNA from muscle tissues of the mandarin fish Siniperca kneri . The cDNA was determined to be of 6987 base pairs in length, encoding a peptide of 1937 amino acids (Genbank accession no. EF446616). A search of encoded protein sequences in the NCBI conserved domain database indicated the presence of all known protein domains for MYH proteins, i.e. the myosin motor domain in the N-terminal region, the DIL domain at the C-terminus, and the ATPase domain. The MYH gene and its protein were expressed predominantly in muscle tissues and weakly in cardiac tissues. Developmentally, the MYH gene was first expressed in the muscle formation stage and continued later on. Our work provided a novel mypsin heavy chain gene sequence in fish biology and the results indicate that the MYH gene and the protein it encodes are important for the growth and development of the mandarin fish, as well as its muscle characterization.     

cDNA cloning and complete primary structures of myosin heavy chains from brushtooth lizardfish Saurida undosquamis and wanieso lizardfish S. wanieso fast skeletal muscles

ABSTRACT:   The complete cDNA sequences encoding predominant types of myosin heavy chain (MYH) in the fast skeletal muscle were determined for brushtooth lizardfish Saurida undosquamis and wanieso lizardfish S. wanieso , which are used as materials for preparing high-quality surimi-based products. The cDNA consisted of 5973 and 5987 bp, respectively, and both encompassed an open reading frame encoding a polypeptide of 1936 amino acid residues. Brushtooth and wanieso lizardfish MYH showed the amino acid sequence identity of 92–93% to white croaker MYH, which was higher than that of 90% to walleye pollack MYH. The putative binding sites for ATP, actin, and regulatory and essential light chains in the subfragment-1 region of brushtooth lizardfish MYH exhibited a high identity with white croaker counterparts as well as the sequences of subfragment-2 and light meromyosin. In contrast, phylogenetic tree, constructed by the neighbor-joining method based on mitochondrial 16S rRNA gene, revealed that the two lizardfish species formed a cluster with walleye pollack, which was paraphyletic with white croaker. Therefore, a good reputation for lizardfish and white croaker to have a high thermal-gel forming ability seemed to be reflected by MYH rather than biological similarity as revealed by the mitochondrial 16S rRNA gene.