罗非鱼碎肉酶法制备蛋白胨的工艺研究

为探讨罗非鱼碎鱼肉酶法制备蛋白胨的加工工艺,采用正交试验的方法,分别研究了木瓜蛋白酶、AS.1398中性蛋白酶、复合蛋白酶水解罗非鱼碎肉制备蛋白胨的工艺条件。结果表明:木瓜蛋白酶的最佳水解工艺条件为pH6.5,温度65℃,水解时间4h,加酶量1250 U/g,蛋白胨得率达12.63%;AS.1398中性蛋白酶最佳水解工艺条件为pH 7.5,温度55℃,水解时间4 h,加酶量750 U/g,蛋白胨得率达13.25%;复合蛋白酶的最佳水解工艺条件为pH 7.0,温度50℃,水解时间4.5 h,加酶量850 U/g,蛋白胨得率达11.43%。     

基于美拉德反应的狭鳕鱼排复配酶解工艺优化

为提高水产品加工副产物中蛋白质资源的综合利用率,采用复合蛋白酶和风味蛋白酶对阿拉斯加狭鳕鱼排进行复配酶解。以水解度为指标,考察初始p H、温度、料液比、加酶量、复配酶比例、时间对酶解液水解度的影响。在单因素试验基础上,采用Box-Behnken中心组合设计和响应面分析法,确定最佳的酶解工艺条件为:初始p H 7.0,温度47℃,料液比1∶2,加酶量3.0%,复合蛋白酶与风味蛋白酶比例1∶1,酶解时间3 h,此时酶解液水解度可达到38.74%。进一步的美拉德反应试验研究表明,在持续酶解9 h后,酶解液水解度、游离氨基酸含量增加趋势均趋于平缓,同时对美拉德反应产物的风味评价差异不显著,因此最终确定酶解时间为9 h。该研究可为利用狭鳕鱼排酶解液进行美拉德反应制备不同肉风味香料提供理论依据。     

鳕鱼下颌肉水解蛋白制备工艺优化

通过单因素实验(蛋白酶种类、加酶量、料液比、水解时间、温度和pH值),对鳕鱼下颌肉进行酶解分析,以水解度(DH)为指标,确定最适蛋白酶为复合风味蛋白酶,其单因子最佳水解条件为:加酶量为1 400 U/g(相对于鱼糜质量)、料液比为1:3、水解时间为5 h、温度为43℃、pH 7.6。经正交试验验证,复合风味蛋白酶的最佳酶解工艺条件为:pH7.0,料液比1∶2,加酶量为1 400 U/g,水解8 h,DH可达42.62%。     

乌贼墨黑色素粗提物的提取工艺优化及其抗氧化能力测定

本文对乌贼墨粉进行超声波辅助蛋白酶水解研究,以水解度为考察指标,筛选最适蛋白酶,并通过单因素试验和L_(16)(4~5)正交试验优化酶解条件。试验结果表明,酸性蛋白酶为墨粉水解的最佳试验用酶,最佳酶解条件为温度50℃、水解时间2 h、加酶量3.0%、反应pH 3.5、料液比1∶15,在此最优条件下对墨粉进行水解试验,制得水解后墨黑色素,并测得水解度为11.45%。水解后墨黑色素氧化产物2,3,5-三羧酸吡咯(PTCA)含量为17.621μg/mg,水解后墨黑色素总抗氧化能力为18.9 U/g,总抗氧化能力保持率为99%。     

壳聚糖固定化胰蛋白酶制备牡蛎肽的研究

以壳聚糖为载体固定化胰蛋白酶,制备牡蛎肽,然后以肽得率为指标,分别研究了pH值,酶加量,酶解时间和温度等单因子对固定化胰蛋白酶酶解牡蛎蛋白质的影响。实验结果表明,酶解牡蛎蛋白的最佳工艺条件是:底物浓度为200μg/mL,加固定化胰蛋白酶量7mg/3mL,酶解时间3h,pH 8.0,温度40℃。在此条件下水解牡蛎蛋白,肽得率可达到20.39%。     

复合酶提取牡蛎抗氧化肽的工艺研究

以牡蛎为原料,首先从木瓜蛋白酶、中性蛋白酶、碱性蛋白酶、胃蛋白酶、胰蛋白酶中筛选出胰蛋白酶为内切酶,以水解度为指标,得到最佳的酶解工艺条件:时间4h,温度50℃,pH8.0,料水比1∶2,加酶量3％(7 500 U/g),水解度为49.50％;同时以水解度为指标,得到外切酶风味蛋白酶的最佳酶解工艺:时间5h,温度50℃,pH8.0,料水比1∶2,加酶量3％ (450 U/g),实际水解度50.95％.最后,以清除羟自由基能力和水解度为指标,探讨内切酶(胰蛋白酶)和外切酶(风味蛋白酶)不同复合酶解方式的抗氧化能力.最终确定,复合酶解制备牡蛎抗氧化肽效果最好,其酶解条件为胰蛋白酶为3％(7 500 U/g)、风味蛋白酶加酶量为3％ (450 U/g),pH8.0,时间5h,料水比1∶2,温度50℃时,水解度高达53.94％,体外清除·OH的EC50为0.56 mg/mL.     

木瓜蛋白酶对罗非鱼下脚料的水解作用

试验结果表明,木瓜蛋白酶水解罗非鱼下脚料蛋白的适宜条件为:底物浓度1∶1.5,温度60℃,加酶量1100 IU/g原料,初始pH,酶解3 h;最佳水解条件下的酶解液氨基态氮含量达1.25 g/L,蛋白质含量5.19%,氮回收率为69.88%。酶活力下降是酶解反应减缓的主要原因之一。     

酶解罗非鱼鱼皮胶制备降血压肽的研究

为探讨罗非鱼鱼皮胶制备降血压肽的酶解工艺,研究选用Neutrase中性蛋白酶、Alcalase碱性内切蛋白酶、胰蛋白酶、木瓜蛋白酶和复合蛋白酶分别对罗非鱼鱼皮胶进行酶解,对其酶解液的水解度进行比较。结果表明复合蛋白酶的酶解效果最好。对复合蛋白酶的酶解工艺进行单因素优化,确定其最佳酶解工艺为:温度50℃、pH值7.0、料液比1∶5、酶量为底物量的1.5%。在此条件下水解6h时,水解度为60%,酶解产物对血管紧张素转化酶(ACE)的抑制率最高,达到78.96%。     

胰蛋白酶水解虾头壳制备钙结合肽的工艺条件优化研究

在单因素试验研究时间、底物浓度、pH,酶浓度和温度对蛋白水解度和酶解液中可溶性钙含量的影响基础上,通过响应面分析法对酶解条件进行了优化,得出最佳酶水解工艺条件为:底物浓度10%,pH 7.4,温度50℃,酶添加量1.39%,水解5h.在此条件下,酶解液水解度为29.0%,可溶性钙含量为176.4 mg/ml,水解液经凝...     

酶法从鳕鱼皮中提取胶原多肽的研究

胶原多肽具有多种生理活性。鳕鱼皮中含有丰富的胶原蛋白。本实验以鳕鱼皮为研究对象，采用酶法从鱼皮中提取胶原多肽，以氨基含量和水解度为指标，通过单因素和正交实验确定酶解的最佳条件。实验结果表明，采用碱性蛋白酶，加酶量4．5％、酶解温度50℃、pH8．5、酶解时间5小时，蛋白质水解度可达到13．87％。     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.     

Cardiac,ventilatory and metabolic responses of two ecologically dissimilar species of fish to waterborne cyanide

Changes in heart rate, ventilatory activity and oxygen consumption were determined in trout (Salmo gairdneri) and brown bullhead catfish (Ictalurus nebulosus) during exposure to a steadily increasing concentration of waterborne cyanide selected to produce death in 8–9 hours for each species. The lethal cyanide concentration for the bullheads was an order of magnitude higher than for trout. Trout developed an immediate and gradually increasing bradycardia throughout the exposure period. Cyanide produced tachycardia in the bullhead followed by a gradual onset of bradycardia as the concentration of cyanide was raised. Pericardial injection of atropine (a muscarinic cholinergic antagonist) indicated that bradycardia in the trout was due initially to increased vagal tone but later due to the direct effect of cyanide on the heart. Hyperventilation in the trout persisted throughout the exposure period, although the rate and amplitude fluctuated and was variable between individual fish. During the last hour of exposure (highest cyanide concentration), ventilation was characterized by rapid, shallow breaths followed by a sudden respiratory arrest. The bullheads exhibited hyperventilation during the first 3 hours of exposure followed by a gradual, linear drop in ventilation rate and amplitude until death occurred. Cardiac and ventilatory responses in both species were attributed to stimulation of central and peripheral chemoreceptors by cyanide. Evidence is presented which suggests the initial response in the bullheads was due, at least in part, to gustatory stimulation by the cyanide. Oxygen consumption of the trout remained above pre-exposure levels for the majority of the test period. Oxygen consumption in the bullhead paralleled the changes in heart and ventilatory rates. Whole-body lactate levels of fingerlings of both species during cyanide exposure were measured to estimate the extent of anaerobiosis. Whole-body lactate levels were much greater in the bullheads than the trout, indicating a higher capacity for anaerobiosis, possibly due to a greater fuel supply. Overall, the trout responded to cyanide in a manner similar to that produced by environmental hypoxia whereas the bullheads experienced a gustatory stimulus which masked the hypoxia-like response.     

An overview of stress physiology of fish transport: changes in water quality as a function of transport duration

This study brings an integrated analysis about the relationship between water deterioration and its physiological consequences in live fish transport. The analysis was focused on the transport water and its deterioration, and physiological challenges imposed on the fish. Usual commercial handling procedures employed to mitigate fish stress during transport were discussed. Future topics of research for the establishment of safer fish transport protocols were proposed. Transport was classified into short (≤8 h) or long transport (>8 h). The main issue in short transports should be the prevention of water pH reduction, while in long transports it is the increase in ammonia. Plasma cortisol is the most employed marker for stress and is acutely elevated upon short episodes of transport, but remains elevated even in long‐transport events. Plasma glucose is perhaps a better marker for handling stress. Plasma lactate, pH, osmolality CO₂ and ions should be more often evaluated. Plasma Na⁺ and Cl^‐ are very useful markers of acidosis, due to their respective exchange for H⁺ and , for acid–base regulation. The establishment of species‐specific transport protocols should be preceded by such combined analyses of water and physiological parameters.     

Are hatchery‐reared abalone naïve of predators? Comparing the behaviours of wild and hatchery‐reared northern abalone,Haliotis kamtschatkana (Jonas, 1845)

Abalone populations have declined worldwide, generating interest in enhancement using hatchery‐reared individuals. In many cases, such restoration efforts have met with limited success due to high predator‐induced mortality rates. Furthermore, the mortality rates of outplanted hatchery abalone are often considerably higher than for wild individuals. This study uses northern abalone (Haliotis kamtschatkana) as a case study to determine whether hatchery‐reared abalone behave differently than their wild counterparts. In the field, outplanted hatchery‐reared abalone were significantly less responsive than wild abalone, in terms of number of abalone responding and intensity of response, to nearby movement and to physical contact with an inert probe. Also, when encountering a cue to which all abalone responded (a seastar predator), hatchery‐reared individuals remained subdued. Anti‐predator behavioural deficits in hatchery‐reared abalone were more pronounced in 4‐year‐old individuals than in 1‐year‐old individuals, suggesting an influence of either age or amount of time spent in the hatchery environment. These behavioural differences are expected to increase the vulnerability of hatchery‐reared abalone to predators, and are likely a major cause of their elevated predator‐induced mortality when outplanted.     

Effects of cadmium on the kinetics of calcium uptake in developing tilapia larvae, Oreochromis mossambicus

The toxic effects of Cd²⁺ on Ca²⁺ influx kinetics in developing tilapia (Oreochromis mossambicus) larvae were evaluated. Addition of 20 µg l^-1 of Cd²⁺ to the environment of 0 and 3 day-old larvae competitively inhibited the Ca²⁺ uptake within 4h resulting in a great increase in K_m values for Ca²⁺ influx (19.3 and 17.4 fold, respectively) as compared with their respective controls. Consequently, the actual Ca²⁺ influx of larvae in solutions of 0.2 mM Ca²⁺ are suppressed by 32–45%. Also, 3 day-old larvae were more sensitive to internally accumulated Cd²⁺ than 0 day-old larvae. Although the Ca²⁺ influx in 0 and 3 day-old larvae may be restored to the levels of their respective controls with 24h of being transferred to a 20 µg l^-1 Cd²⁺ solution, total body Ca²⁺ content was significantly reduced in 3 day-old larvae. Increased Ca²⁺ uptake efficiency ensures sufficient Ca²⁺ for normal growth. However, rapid increase in Ca²⁺ influx after hatching also leads to higher Cd²⁺ uptake. Exposure to Cd²⁺ will lead to a drop in body Ca²⁺ content resulting in retardation of larval growth. Therefore, we conclude that if Ca²⁺ uptake is interfered with at this critical stage of development, larvae will not be able to maintain normal levels of body Ca²⁺ and will show signs of Cd²⁺ poisoning.     

Migratory dynamics of stream-spawning longnose gar (Lepisosteus osseus)

Abstract– Literature evidence suggests that lake-dwelling longnose gar (Lepisosteus osseus) enter tributary streams to spawn, Until the present study, the dynamics of this breeding migration had never been investigated quantitatively. During the summers of 1991 and 1992, longnose gar were captured as they entered Weaubleau Creek, Missouri, a tributary of Harry S. Truman Reservoir. The in-stream spawning migration began in early April and ended in late May, and was positively correlated with stream flow and water level, and negatively correlated with water temperature. In-stream residence times ranged from 15 to 94 days, with males exhibiting longer residence times than females. Once in-stream, longnose gar travelled as far as 10 km upstream and occupied certain pools at greater relative frequencies. Although the reason for this preferential utilization is not completely understood, it may relate to pool depth and riffle proximity. Longnose gar disperse from the spawning stream great distances, with gar captured in Weaubleau Creek being recaptured up to 48 km away. This information should provide fisheries biologists the means to consider the reproductive ecology of this species in their conservation and management decisions.     

Nutritional regulation of hepatocyte fatty acid desaturation and polyunsaturated fatty acid composition in zebrafish (Danio rerio) and tilapia (Oreochromis niloticus)

The desaturation and elongation of [1-¹⁴C]18:3n-3 was investigated in hepatocytes of the tropical warm freshwater species, zebrafish (Danio rerio) and Nile tilapia (Oreochromis niloticus). The hepatocyte fatty acid desaturation/elongation pathway was assayed before and after the fish were fed two experimental diets, a control diet containing fish oil (FO) and a diet containing vegetable oil (VO; a blend of olive, linseed and high oleic acid sunflower oils) for 10 weeks. The VO diet was formulated to provide 1% each of 18:2n-6 and 18:3n-3, and so satisfy the possible EFA requirements of zebrafish and tilapia. At the end of the dietary trial, the lipid and fatty acid composition was determined in whole zebrafish, and liver, white muscle and brain of tilapia. Both zebrafish and tilapia expressed a hepatocyte fatty acid desaturation/elongation pattern consistent with them being freshwater and planktonivorous fish. The data also showed that hepatic fatty acid desaturation/elongation was nutritionally regulated with the activities being higher in fish fed the VO diet compared to fish fed the FO diet. In zebrafish, the main effect of the VO diet was increased fatty acid Δ6 desaturase activity resulting in the production of significantly more 18:4n-3 compared to fish fed the FO diet. In tilapia, all activities in the pathway were greater in fish fed the VO diet resulting in increased amounts of all fatty acids in the pathway, but primarily eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). However, the fatty acid compositional data indicated that despite increased activity, desaturation of 18:3n-3 was insufficient to maintain tissue proportions of EPA and DHA in fish fed the VO diet at the same level as in fish fed the FO diet. Practically, these results indicate that manipulation of tilapia diets in commercial culture in response to the declining global fish oil market would have important consequences for fish fatty acid composition and the health of consumers. Scientifically, zebrafish and tilapia, both the subject of active genome mapping projects, could be useful models for studies of lipid and fatty acid metabolism at a molecular biological and genetic level. This revised version was published online in August 2006 with corrections to the Cover Date.     

Production of diatom–bacteria biofilm isolated from Seriola lalandi cultures for aquaculture application

Controlled incorporation of selected microalgae and bacteria in aquaculture systems can be beneficial because they can act as microbiological control. That is why the characteristics of biofilm generated naturally in Seriola lalandi culture cages were analysed, their potential benefit to the growth of larvae was studied, and their controlled use for improving the larval viability and as a vector to improve incorporation of previously studied probiotic bacteria was tested. According to biodiversity results, these biofilms are composed of a diatom–bacteria mix showing a decrease in biodiversity in laboratory culture conditions being dominated by Navicula phyllepta and bacteria of the family Rhodobacteraceae. This can be produced on mesh substrates incorporated in bioreactors with rapid growth rate and adhesiveness. Preliminary results from the addition of substrates with this specific biofilm in larvae culture systems showed that it is consumed by the larvae without negative effects, while positive effects on the viability of larvae in combination with probiotics were observed. Considering preliminary results, the addition of these specific substrates with diatom–bacteria biofilms could be a good improvement for aquaculture systems and together with the use of probiotics can contribute to maintaining a stable and controlled system improving the viability of the larval fish culture in its early stages.