Manipulation of fatty acid composition of Nile tilapia (Oreochromis niloticus) fillets with flaxseed oil

Nile tilapia (Oreochromis niloticus) is one of the most consumed species among freshwater fish reared in Brazil. However, studies show low levels of n‐3 fatty acids in freshwater fish reared in captivity in comparison with those reared in their natural habitats. The Nile tilapia used in this study were raised in captivity for a period of 5 months and fed varying amounts (0, 1.25, 2.5, 3.75 and 5%) of flaxseed oil as a substitute for sunflower oil (control). No significant differences (P > 0.05) in total lipid (TL) content were found between fillets of tilapia fed the different diets. TL analysis of fatty acid methyl esters by capillary gas chromatography revealed a total of 50 components common to all treatments studied. The major fatty acids present were linoleic acid (18:2n‐6), oleic acid (18:1n‐9) and palmitic acid (16:0). All treatments led to significant % increases in α‐linolenic acid (18:3n‐3), eicosapentaenoic acid (20:5n‐3) and docosahexaenoic acid (22:6n‐3). Increases in both total n‐3 fatty acids and total polyunsaturated fatty acids were observed concomitantly with decreases in total n‐6 fatty acids, resulting in increases in n‐3/n‐6 ratio, with increasing level of flaxseed oil in the feed. Thus feed supplementation with flaxseed oil contributed greatly to raising the nutritional lipid value of Nile tilapia fillets. Copyright © 2007 Society of Chemical Industry     

利用罗非鱼卵黄原蛋白建立雌激素污染的检测方法

目的以尼罗罗非鱼为实验对象,以卵黄原蛋白(Vg)为生物标志物开发环境雌激素的生物检测技术。方法采用凝胶过滤与离子交换层析相结合的方法从17β-雌二醇诱导后的罗非鱼血浆中分离纯化Vg,对纯化的蛋白进行鉴定后制备多克隆抗血清,建立Vg的酶联免疫吸附测定实验(ELISA),并用于样品测定。结果建立的ELISA工作范围为15.6~1000 ng/m L,组内与组间差异分别为6.85%与6.79%,能有效检测100、500、1000μg/L双酚A对雄鱼血浆Vg的诱导效应。结论本研究建立的Vg ELISA具有较高的敏感度、特异性与精确度,可为水体环境雌激素污染检测提供重要工具。     

Selenium fractionation from plasma, muscle and liver of Nile tilapia (Oreochromis niloticus)

This paper presents the results obtained from selenium fractionation in plasma, muscle and liver samples of Nile tilapia’s (Oreochromis niloticus) after protein separation. The plasma, muscle and liver proteome was obtained by 2D-PAGE, and selenium in protein spots was qualitatively and quantitatively determined by synchrotron radiation X-ray fluorescence and graphite furnace atomic absorption spectrometry (GFAAS). The fluorescence spectra indicated the presence of selenium in three protein spots of plasma, two of muscle and one of liver. Selenium was found to be distributed mainly in proteins with a molar mass smaller than 57.0 kDa and with pI in the range of 5.9–9.6, with one exception in the plasma sample, which presented protein with a molar mass of 60.0 kDa. After acid mineralization of the protein spots, a GFAAS determination of the concentration of selenium bound to these proteins indicated a range of 1.35–6.82 mg per g of protein.     

Effect of chitosan–Jicama starch coating on changes in qualities of fresh Nile tilapia (Oreochromis niloticus) fillets during ice storage

The effect of chitosan (0.5%)/Jicama starch (0%–4%)‐based edible coating on the quality of Nile tilapia (Oreochromis niloticus) fillets was evaluated over ice storage time. All samples were periodically analysed for pH value, thiobarbituric acid (TBA), total volatile basic nitrogen (TVB‐N), electrical conductivity (EC), total viable counts (TVC), total psychrotrophic counts (TPC), drip loss, colour, hardness and sensory characteristics. Results demonstrated that the quality of Nile tilapia fillets was preserved by the film containing chitosan and/or Jicama starch. Compared with chitosan coating alone (0.5% chitosan/0.25% glycerol) (P < 0.05), T3 (0.5% chitosan/1% Jicama starch/0.25% glycerol) had a better effect on the drip loss, TBA, TVC, TPC, hardness and sensory characteristics of the samples, thus indicating that low Jicama starch concentration (1%) enriched the coating ability of chitosan in extending the shelf life of Nile tilapia fillets.     

不同脂肪酸甲酯化方法对共轭亚油酸分析的影响

共轭亚油酸(CLA)是亚油酸的位置和几何异构体,因其具有多种生物学功能而成为人们关注的焦点。气相色谱法是分析CLA的一种简便而有效的方法,通过分析可以确定CLA的组成和含量。气相色谱法分析CIA先涉及脂肪酸的甲酯化,脂肪酸的甲酯化方法可分为3大类,酸催化、碱催化和三甲基硅重氮甲烷(TMS)法。一般游离型脂肪酸的甲酯化可采用酸催化或TMS法,而三甘油酯型的脂肪酸可采用酸催化或碱催化法。主要探讨3种甲酯化方法在不同结构的CLA气相色谱分析中的异同,通过薄层层析(TLC)和气相色谱(GC)测定,发现甲酯化过程中脂肪酸酯化的程度各不相同,CLA甲酯化后组成发生了异构化。结果表明,CLA经过酸催化法后得脂肪酸含量为73.34%,而TMS法为82.47%;酸催化法后反反CLA(tt-CLA)含量为24.66%。     

Chemoenzymatic synthesis of structured triacylglycerols with conjugated linoleic acids (CLA) in central position

A chemoenzymatic process for the production of structured triacylglycerols (TAG) containing CLA at sn2 position and lauric acid at external ones is proposed. First, castor bean oil was chemically dehydrated and isomerised to obtain a new modified oil with very high proportion of CLA (>95%). Then, this new oil was used for enzymatic transesterification allowing the grafting of lauric acid at external positions of the TAG backbone by using 1,3 regioselective enzymes. Among these, Aspergillus niger lipase was not satisfactory giving very low lauroyl incorporation (<5%) On the contrary, lipases from Thermomyces lanuginosa (Lipozyme TL IM) and from Carica papaya latex allowed good reaction yields. The effect of the type of acyl donor was studied. With alkyl esters T. lanuginosa lipase provided a final incorporation of 58.9% after 72 h corresponding to 88.4% transesterification yield. Concerning C. papaya lipase, incorporation of lauroyl residues was lower than Lipozyme TL IM. This lipase exhibited higher performance with lauric acid accounting for 44.7% lauroyl incorporation at the end of reaction for a 67.1% transesterification yield. The effect of the substrates mole ratio was also evaluated. It was observed that a 1:3 TAG/acyl donor mole ratio was the most efficient for both lipases. Finally, fatty acids regiodistribution of the newly formed structured TAG was determined. With Lipozyme TL IM, the proportion of lauric acid incorporated at the sn2 position did not exceed 5.4% after 72 h while with C. papaya lipase a more pronounced incorporation of lauroyl residues at the central position (8.8%) was observed.     

Original article: Optimisation of extraction conditions and characteristics of skin gelatin from Nile tilapia (Oreochromis niloticus)

Response surface methodology was used to optimise gelatin extraction conditions from the skin of Nile tilapia (Oreochromis niloticus), and characteristics of the gelatin were determined. Concentration of NaOH (%, X₁), alkaline treatment time (h, X₂), concentration of HCl (‰, X₃) and acid treatment time (min, X₄) were chosen for independent variables. Dependent variable was yield of gelatin (%, Y). Optimal conditions were X₁ = 3.2 (%), X₂ = 2.3 (h), X₃ = 0.7 (‰) and X₄ = 84 (min), and predicted value of response optimal conditions was Y = 20.4%. Actual value was 19.3% by verification experiments under optimal conditions. Crude protein content of the tilapia skin gelatin was 88.5%. The content of imino acids including proline and hydroxyproline in the gelatin was 185 residues per 1000 total amino acid residues. Its gel strength was 260 g. The gelling and melting points were 18.0 and 22.4 °C, respectively.     

Calcium-binding peptides derived from tilapia (Oreochromis niloticus) protein hydrolysate

Calcium-binding peptide was derived from protein hydrolysates. In this study, tilapia protein at a concentration of 2 % (w/v) was hydrolyzed using various proteases including Alcalase 2.4 L, Flavourzyme 1,000L, Protease GN, and papain at 50 °C, pH 8 for 6 h. It was found that the degree of hydrolysis increased with the time of the incubation in all cases. The highest calcium-binding capacity of the hydrolysate was 65 mg/g protein at 27.7 % degree of hydrolysis by Alcalase 2.4 L. The molecular weight of the calcium-binding peptides characterized by gel-filtration chromatography on a Sephadex G-25 was 1.2 kDa. The calcium-binding motif of the hydrolyzed peptides identified by the automated Edman degradation was a short peptide (Trp-Glu-Trp-Leu-His-Tyr-Trp). The results of this study suggested that tilapia protein is a good source for calcium-binding peptides.     

Feasibility of fishmeal replacement by shrimp head silage protein hydrolysate in Nile tilapia (Oreochromis niloticus L) diets

This study provides information on the use of shrimp head silage protein hydrolysate (SPH) as an alternative protein source for tilapia feeding. Six diets (28% protein, 12% lipid) were prepared where fishmeal protein was replaced at levels of 0 (control), 10, 15, 20, 25 and 30% with the hydrolysate. The diets were supplied to Nile tilapia fry (338 mg initial weight) stocked in plastic recirculating 20 l tanks (10 animals per tank), with three replicates per treatment. After an 8 week experimental period, fish fed the diets containing 10 and 15% SPH showed significantly better performance in terms of final body weight, weight gain (%), mean daily weight gain (mg day^?1), specific growth ratio and feed conversion ratio than those fed the control diet (fishmeal as protein source) and higher‐SPH diets. It is concluded that shrimp head hydrolysate is a promising alternative protein source for tilapia feeding, improving growth ratio at dietary inclusion levels as high as 15%. In addition, the diets with added shrimp silage protein were well accepted by the fish, which avidly consumed the feed during the experiment. © 2002 Society of Chemical Industry     

Production of conjugated fatty acids by lactic acid bacteria

Conjugated fatty acids have attracted much attention as a novel type of biologically beneficial functional lipid. Some isomers of conjugated linoleic acid (CLA) reduce carcinogenesis, atherosclerosis, and body fat. Considering the use of CLA for medicinal and nutraceutical purposes, a safe isomer-selective process is required. The introduction of biological reactions for CLA production could be an answer. We screened microbial reactions useful for CLA production, and found several unique reactions in lactic acid bacteria. Lactic acid bacteria produced CLA from linoleic acid. The produced CLA comprised a mixture of cis-9,trans-11-octadecadienoic acid (18:2) and trans-9,trans-11-18:2. Lactobacillus plantarum AKU 1009a was selected as a potential CLA producer. Using washed cells of L. plantarum AKU 1009a as a catalyst, CLA production from linoleic acid reached 40 mg/ml under the optimized conditions. The CLA-producing reaction was found to consist of two successive reactions, i.e., hydration of linoleic acid to 10-hydroxy-12-octadecenoic acid and dehydrating isomerization of the hydroxy fatty acid to CLA. On the basis of these results, the transformation of hydroxy fatty acids by lactic acid bacteria was investigated. Lactic acid bacteria transformed ricinoleic acid (12-hydroxy-cis-9-octadecenoic acid) to CLA (a mixture of cis-9,trans-11-18:2 and trans-9,trans-11-18:2). Castor oil, which is rich in the triacylglycerol form of ricinoleic acid, was also found to act as a substrate for CLA production by lactic acid bacteria with the aid of lipase-catalyzed triacylglycerol hydrolysis. L. plantarum AKU 1009a produced conjugated trienoic fatty acids from alpha- and gamma-linolenic acid. The trienoic fatty acids produced from alpha-linolenic acid were identified as cis-9,trans-11,cis-15-octadecatrienoic acid (18:3) and trans-9,trans-11,cis-15-18:3. Those produced from gamma-linolenic were cis-6,cis-9,trans-11-18:3 and cis-6,trans-9,trans-11-18:3. The conjugated trienoic fatty acids produced from alpha- and gamma-linolenic acid were further saturated by L. plantarum AKU 1009a to trans-10,cis-15-18:2 and cis-6,trans-10-18:2, respectively.     

Development of a Nile tilapia (Oreochromis niloticus) gut microbiota-derived bacterial consortium with antibacterial activity against fish pathogens

Multistrain potential probiotic mixtures were assembled from isolates from Nile tilapia (O. niloticus) gut microbiota. Lactococcus lactis A12, Priestia megaterium M4, and Priestia sp. M10 demonstrating antibacterial activity against two fish pathogens, Streptococcus agalactiae and Aeromonas hydrophila, were proportion optimized using mixture design software. Two potential probiotic mixtures were chosen and compared for storage viability: one containing 61% of strain A12, 16% of M4, and 23% of M10; the other containing 28% of M4 and 72% of M10. The mixtures were combined with maltodextrin or fish feed, freeze-dried, and bacterial counts were monitored over 28 days while stored at 4 °C or 25 °C. At 4 °C storage, the multistrain potential probiotics were stable in maltodextrin and fish feed. In conclusion, a mixture design may be used to define a multistrain potential probiotic, and freeze-drying while combined with maltodextrin or fish feed followed by refrigeration could be employed to stabilize these tilapia-derived potential probiotics.     

Enzymatic hydrolysis of Nile tilapia (Oreochromus niloticus) myofibrillar proteins: effects on nutritional and hydrophilic properties

Protein concentrate (PC) was prepared from eviscerated and mechanically deboned fish. The pulp was washed sequentially with 0.05 M NaCl and 4 g l^?1 NaHCO₃ solutions, with cold ethanol and then with hot (50 °C) ethanol. Subsequently, it was dried (40 °C) and ground to 60 mesh. Enzyme (Flavourzime^®) hydrolysates were prepared with a degree of hydrolysis (DH) ranging from 3.5 to 45%. The PC was characterized for proximate composition, amino acid profile, physical and functional properties. The influence of DH on functional and nutritional properties was evaluated. Hygroscopicity increased in all hydrolysates for environmental relative humidities above 40% and solubility increased rapidly for DH above 7%. Water retention, water absorption and oil absorption all decreased as a function of DH. The pH (3 to 9) influenced the water and oil absorption of the hydrolysate with 7% DH. A pH near the isoelectric point tends to decrease water absorption but increase oil absorption capacity. The insoluble fraction (IF) of the hydrolysates presented a better amino acid profile than the soluble fraction (SF). The nutritional indexes determined for the IF did not differ from the total hydrolysate, but they were consistently higher than those for the SF. Copyright © 2003 Society of Chemical Industry     

Study on the electric conduction properties of fresh and frozen–thawed grass carp (Ctenopharyngodon idellus) and tilapia(Oreochromis niloticus)

Electric conduction properties of fresh and frozen–thawed grass carp (Ctenopharyngodon idellus) and tilapia (Oreochromis niloticus) were studied by measuring their impedances under different frequencies. The impedances varied as frequency changed. Although impedances of both fresh and frozen–thawed fish decreased as frequency increased from 1 to 20 kHz, change ratio of impedance (Q value) of fresh fish was evidently higher than that of frozen–thawed fish. The Q values of fresh grass carp and tilapia on the 1st, 3rd, 6th, 8th, 10th days were 60.53%, 52.24%, 35.47%, 26.86%, 21.17% and 73.13%, 52.70%, 41.06%, 39.28%, 36.31%, while frozen–thawed grass carp and tilapia were 9.65%, 11.21%, 9.58%, 8.89%, 9.13% and 18.26%, 12.80%, 13.46%, 12.16%, 12.91%, respectively. Electric conduction is a rapid, easy to use, portable and cost‐effective method for in situ analysis at practical level. Therefore, it is feasible to identify fresh and frozen–thawed fish according to the change ratio of their impedances.     

罗非鱼鱼肉及组分蛋白酶解产物的抗氧化特性

采用胃蛋白酶、中性蛋白酶、Alcalase2.4L碱性蛋白酶分别酶解罗非鱼肉蛋白,研究比较不同蛋白酶在1、2、4、6、8h对罗非鱼肉蛋白的酶解效果及产物的抗氧化活性,结果表明:3种酶的酶解产物都具有一定的抗氧化活性,中性蛋白酶酶解产物的水解度和蛋白回收率最高,胃蛋白酶酶解产物对DPPH自由基和羟自由基的清除效果最好;综合各项指标选择中性蛋白酶对罗非鱼肉的组分蛋白(肌原纤维蛋白、肌浆蛋白、基质蛋白)进行4h酶解,研究比较不同组分蛋白的酶解效果及产物的抗氧化活性,结果表明:组分蛋白中基质蛋白酶解产物的蛋白回收率最高,为89.8%,肌原纤维蛋白酶解产物的水解度最高,为10.1%,肌浆蛋白酶解产物的抗氧化活性最好,羟自由基和DPPH自由基的IC50值分别为12.352mg/m L和3.554mg/m L。