963 resultados para glutathione peroxide


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水稻既是我国三大粮食作物之一,又是基因组学研究的模式材料,在生产实践和科学研究中都占有极其重要的地位。基因组学研究取得的巨大成就以前所未有的深度和广度推动了生命科学各个研究领域的飞速发展。水稻基因组的破译是水稻科学研究的重要里程碑,同时也宣告了功能基因组学时代的到来。蛋白质组学是研究细胞内全部蛋白质的动态表达及其相互关系的新兴学科,是功能基因组学研究的重要组成部分和战略制高点。 本论文采用高分辨率的蛋白质双向电泳分离技术和高通量的蛋白质质谱分析技术以及生物信息学等手段,开展水稻灌浆期茎蛋白质组表达模式和水稻幼苗脱黄化过程的比较蛋白质组学研究,探讨茎生长发育规律和水稻应答光信号相关蛋白质及其网络调控机制,是学科前沿与实际应用的有机结合,在科研和生产实践中都具有重要的意义。 首先,分别构建了灌浆期水稻顶端茎段和水稻黄化幼苗的蛋白质组表达谱。并对其中185个目的蛋白点进行了MALDI-TOF/MS分析和数据库检索鉴定。共有149个蛋白质得到了鉴定,蛋白质鉴定的成功率为80.5%。这些被鉴定的蛋白质分属118个基因的表达产物,根据它们功能可以分为13种不同的类别,其中绝大多数为能量产生和代谢以及抗性相关的蛋白质。 在水稻灌浆期顶端茎段表达的蛋白质中,与能量和物质代谢相关的蛋白质例如ATPase、磷酸丙糖异构酶,6-磷酸葡萄糖异构酶等占有很高比例,说明茎段组织中具有很强的代谢活动。与生长发育相关的蛋白质包括beta-tubulins、无机焦磷酸酶(inorganic pyrophosphatase)、液泡质子ATP酶(vacuolar proton-ATPase)以及UDP葡萄糖焦磷酸酶等的大量累积,显示出顶端茎段细胞分裂和生长迅速;同时,贮存多糖和结构多糖也在旺盛合成。G蛋白、GDP释放抑制因子等信号传导蛋白以及苯丙氨酸氨解酶、谷胱苷肽S转移酶(glutathione S-transferase,GST)、抗坏血酸过氧化酶(ascorbate peroxidase,APX)以及超氧化物歧化酶(superoxide dismutase,SOD)等抗性相关蛋白质在该时期丰度表达,表明在灌浆期水稻顶端茎段能够迅速感受并传递外界信号,从而使得其在遭受胁迫时能够立刻启动抗逆防御系统,最大限度地降低不利环境对种子发育的影响。 在黑暗中萌发和生长的水稻黄化幼苗随着光照时间(0~24小时)的延长,能通过双向电泳后检测到的蛋白质逐渐变少,24小时后趋于稳定,相当于正常光照条件下生长的水稻幼苗蛋白质组表达谱。进一步分析表明,在黄化苗中,分解代谢及能量产生相关的蛋白如丙糖磷酸异构酶、琥珀酰辅酶A连接酶、异戊酰辅酶A脱氢酶与ATPase等的表达量比较丰富;另外,还可能启动了脂肪酸的α氧化分解途径,以供黑暗中生长所需的物质和能量。当黄化幼苗光照后,与光合作用及物质合成相关的一些蛋白质表达量增加,而那些分解代谢相关酶类则有所下降。同时,鸟核苷酸结合蛋白β亚基类似蛋白、20S proteasome以及Bowman Birk trypsin inhibitor等信号传递及抗性相关蛋白随着光照时间的延长而减少,说明黑暗胁迫条件下水稻幼苗启动了相关的抗逆途径。叶绿素合成途径中的蛋白酶胆色素原脱氨基酶和金属鳌合酶在脱黄化过程中表达量有所下降,可能是因为叶绿素合成产物具有反馈抑制作用。 本研究首次利用蛋白质组学方法来解析水稻灌浆期茎蛋白质组表达模式和水稻黄化幼苗响应光因子的蛋白质组变化情况,鉴定了一些有价值的蛋白质,并得到了它们的表达特点和相关数据,为更好地理解水稻顶端茎秆的生长特点和功效、水稻应答黑暗胁迫和光形态建成以及光合作用机理等提供了分子证据。

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磷缺乏已成为制约世界农业生产的重要因子。植物根系的大小和形态是决定植物吸收土壤磷能力的重要因素,而且根系的生长发育与磷素的分布及其有效性密切相关。关于磷酸盐调节植物根系生长研究已有很多报道,但其生理和分子机制仍不清楚。一氧化氮 (NO) 是一种重要的气体信号分子,参与调控植物的生长发育和对多种逆境胁迫的应答反应。本文选用拟南芥为实验材料,研究探讨了NO与缺磷诱导的拟南芥根系形态变化之间的关系,主要结果如下: 用正常磷水平 (1 mM) 和低磷水平 (1 µM) 处理拟南芥幼苗,发现低磷抑制主根伸长,刺激侧根发生。外源NO供体销普纳 (SNP) 也抑制主根、刺激侧根生长,与低磷诱导根系形态变化相似。NO清除剂c-PTIO和一氧化氮合成酶 (NOS)抑制剂L-NNA均可部分减缓由低磷引起的对主根生长的抑制和对侧根的刺激作用。暗示低磷诱导的拟南芥根系形态的变化可能与NO含量的降低有关。 利用NO荧光标记物DAF-FM和激光共聚焦显微成像技术,本研究发现缺磷6 h和24 h后根细胞内源NO含量显著增加,而且NOS 抑制剂能减少低磷诱导的根细胞NO含量的增加。与正常供磷处理相比,低磷处理6 h和24 h,拟南芥根中编码与NO合成相关的基因(AtNOA1)的表达量增加,缺磷24 h后根中NOS酶活性升高。为了明确低磷诱导的NO 增加是否与硝酸还原酶(NR)介导的NO合成有关,本论文进一步研究了低磷对拟南芥硝酸还原酶活性和编码NR基因 (AtNR1和AtNR2)表达的影响。研究发现低磷处理6 h和24 h后和AtNR1和AtNR2基因的表达均没有变化,且蛭石中生长的拟南芥缺磷1个月后NR活性也没有发生变化;拟南芥的NR双突变体nia1,nia2在低磷处理24 h后,其根中的内源NO含量表现出与野生型相同的增加。因此这些研究结果表明,缺磷后拟南芥根细胞NO的含量增加主要由于NOS的活性升高,而与NR介导的NO合成无关。 已有资料表明低磷诱导植物根细胞内源过氧化氢(H2O2)分布和含量的变化。本论文研究了低磷处理对用H2O2标记物CM-H2DCFDA标记不同磷处理下的拟南芥根中的H2O2。研究发现,缺磷6 h根中H2O2的分布无明显变化,缺磷24 h后H2O2呈斑块状分布,且多集中在根尖伸长区。缺磷24 h后,叶片中的抗氧化保护酶—超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性没有明显变化。说明缺磷24 h 后产生的H2O2没有引起氧化胁迫,而是作为一种信号分子,与NO相互作用共同介导低磷胁迫的应答反应。关于NO与H2O2在低磷诱导的根形态变化中的信号转导过程还有待进一步研究。

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谷胱甘肽还原酶(GR,EC1.6.4.2)是一重要的抗氧化酶,许多生理学和遗传工程研究都证明GR酶在抗氧化中的重要作用。但改变GR酶怎样影响植物的抗氧化系统却不清楚。GR是抗坏血酸-谷胱甘肽循环途径中的重要组成部分,其功能必然与其密切相关。本文用RNAi技术获得具有较低GR酶活性的转基因烟草,系统测定了非胁迫条件和胁迫条件下抗坏血酸-谷胱甘肽循环的变化,得出以下主要结果: 1.选择一烟草叶绿体GR酶编码基因(X76293, gi: 431954)进行RNAi载体构建,构建好的双元载体转化根癌农杆菌LBA4404,然后侵染转化烟草叶圆片。获得的转基因烟草具有30-70%的GR酶活性。分子检测结果表明GR在RNA和蛋白水平上与GR酶活性的变化一致。文中我们第一次用2-D电泳对烟草中GR同工酶进行分析,并确定发生抑制的GR同工酶在细胞中的定位。2-D电泳后的Western杂交检测到烟草的10种GR同工酶,pI值分布在4.5-6.3,其中3种GR同工酶定位在叶绿体内,其蛋白量占据所有GR酶含量的大部分。RNAi发生在叶绿体内和叶绿体外,表明发生抑制的GR同工酶的基因序列具有很高的同源性。igr转基因烟草在表型上与野生型对照烟草无明显差异。 2.所有igr转基因植株和对照植株中的活性氧(O2-和H2O2)、MDA含量和光合作用都无明显差异,表明正常生长条件下GR酶活性的降低不会引起氧化胁迫。测定正常生长条件下igr转基因烟草中谷胱甘肽库的变化。结果表明与对照烟草相比,GR酶活性降低70%会引起转基因植株中GSH/GSSG比率明显降低,而GSH和GSSG的含量稍有增加;测定抗坏血酸-谷胱甘肽循环的变化,结果显示igr转基因烟草中DHAR和MDHAR的酶活性升高,表明非胁迫条件下较低的GR酶活性可能会诱导抗坏血酸-谷胱甘肽循环不能正常的运转。这一作用可能与改变的谷胱甘肽库有关。GR酶活性降低30%的转基因烟草中未检测到这些变化,表明70%的GR酶活对于非胁迫条件下igr转基因烟草可能是足够的。 3. MV处理结果显示,igr转基因烟草的离体叶圆片和活体植株在MV处理后都发生比对照烟草严重的光漂白作用。igr转基因烟草的活性氧和MDA含量明显高于对照烟草,igr转基因烟草的光合作用明显低于对照烟草。以上这些指标表明igr转基因烟草对MV处理更为敏感。MV处理条件下igr转基因烟草谷胱甘肽的含量明显高于对照烟草,但是GSH/GSSG的比率明显低于对照烟草,GR酶活性仍明显低于对照烟草,表明在MV胁迫条件下igr转基因烟草中较低的GR酶活性不能有效的将GSSG还原生成GSH。igr转基因烟草中较高的谷胱甘肽净含量说明其谷胱甘肽的合成能力提高,但这仍不能补偿胁迫条件下较低GR酶引起的GSH/GSSG比率降低。MV处理条件下igr转基因烟草和对照烟草相比ASC的含量大大降低,导致DHA/ASC明显升高。测定MDHAR和DHAR的结果表明,MV处理后igr转基因烟草的MDHAR酶活性明显降低,这表明较低的GR酶活性引起ASC再生循环受到抑制。MV处理后较低的GR酶还引起igr转基因烟草中APX的活性大大降低。以上这些结果表明MV处理条件下降低GR酶活性会削弱抗坏血酸-谷胱甘肽循环,从而引起活性氧的大量积累,造成严重的氧化伤害。 4.低温处理的结果和MV处理的结果稍有不同。在GR酶活性较高的i2转基因烟草中所有检测指标与对照烟草无明显差异。而GR酶活性较低的i21、i28和i42植株与对照烟草相比表现出明显差异。低温下生长的对照烟草叶绿素含量明显高于i21、i28和i42植株。i21、i28和i42中活性氧(O2-和H2O2)和MDA的含量都明显高于对照烟草,表明低温处理下i21、i28和i42受到更严重的胁迫伤害。与MV处理后的变化相似,低温处理后i21、i28和i42中较低的 GR酶活性导致GSH/GSSG大大降低,ASC再生循环受抑制,APX活性明显降低,从而使抗坏血酸-谷胱甘肽循环不能高效的清除活性氧,导致ROS和MDA的大量积累,造成严重的低温伤害。

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近年来大量研究表明水杨酸(salicylic acid, SA)在植物抵抗生物胁迫与非生物胁迫中都发挥着重要作用。然而在一些单子叶植物如水稻中SA的作用迄今仍不是很清楚。为了更深入地了解SA在水稻抵御冷胁迫中的作用,本研究选用两个抗冷性不同的水稻品种:‘长白九’(Oryza sativa cv. ‘Changbaijiu’)和‘中鉴’(Oryza sativa cv. ‘Zhongjian’)作为实验材料,其中‘长白九’为抗冷性较强的品种,而‘中鉴’为冷敏感的品种。在水稻幼苗长至三叶期后,分别对其施以三种浓度(0.5 mM, 1.0 mM, 2.0 mM)的SA溶液预处理24 h,然后置于5 °C下进行冷处理24 h。形态学观察及各项指标的测定结果表明: 一、冷处理后,‘长白九’和‘中鉴’根与叶片中的SA含量都大幅提高,且结合态SA升高的幅度明显大于其自由态形式。 二、外施不同浓度的SA溶液于水稻根部,24 h后,大量SA尤其是结合态SA积累于根中,且其积累量与处理浓度成正相关;而叶片中积累的SA则较少。 三、形态学及生理指标的测定结果显示,SA预处理没有提高甚至降低了两个水稻品种幼苗的抗冷性。并且SA处理浓度越大,幼苗受到冷伤害程度的越高。 四、对水稻幼苗叶片与根中的抗氧化酶活性进行分析发现,常温下SA处理显著提高了‘长白九’和‘中鉴’根中过氧化氢酶(catalase, CAT)和谷胱甘肽还原酶(glutathione reductase, GR)的活性;而在低温下SA预处理反而降低了两种水稻叶片与根中部分抗氧化酶的活性,推测低温下抗氧化酶活性的下降可能与水稻幼苗抗冷性的降低有关。 五、尽管两个水稻品种具有不同的冷敏感性,然而外施水杨酸均加剧了其低温伤害。分析认为,外施水杨酸后,水稻根部大幅升高的内源SA水平可能加剧了活性氧的产生,破坏了植物细胞内部的氧化还原平衡,从而导致水稻幼苗受到的冷害加重。

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Shark livers are considered as an important raw material providing a quality fish oil. It has been reported to aid white — blood-cell production and act as an active ingredient in hemorrhoid treatments. It is also reported that liver oil as a good supplement of vitamin A and poly-unsaturated fatty acids which are important to the development of brain cells in human. Freshness of livers is very important to extract better quality oil. In Sri Lanka, the annual shark production amounts to 8000t, however the quality of livers collected from landing sites has not being measured yet. Present study was conducted to evaluate the quality of silky (Charcarninus fakiformis) shark livers available in Negombo and Beruwala landing sites in the West Coast of Sri Lanka and also to study the relationship between organoleptic and bio-chemical correlation on freshness of shark livers. Liver samples which were collected from landing sites in the West coast of Sri Lanka, were evaluated for external and internal colour, texture and odour. Total volatile nitrogen (TVN), pH value, free fatty acid (FFA%) and peroxide (PV) values of livers were also determined to assess quality. According to the organoleptic scoring system 4.3% of liver samples were categorized as best in quality while 30.4%, 56.5% and 8.7% rated as good, medium and poor in quality respectively at the Negombo and Beruwala landing sites. Bio-chemical analysis showed that the better quality livers had the highest score for sensory evaluation and low values for TVN, FFA and peroxide value while low quality livers gave low score for sensory evaluation and high TVN, FFA, peroxide values. Correlation coefficient of organoleptic scores against total volatile nitrogen value, pH value, free fatty acid % and peroxide value of shark livers were determined by statistical analysis. Organoleptic score of shark livers was found to be highly.

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The overall quality of five SIS products was found in good condition up to 2 months storage on the basis of organoleptic, biochemical and bacteriological characteristics and all the products was excellent in sealed packed condition up to 45 days of storage. However, quality of the products stored in open air atmospheric temperature was found excellent for first 15 days. In an average the initial moisture content was in the range of 13.5 to 15.0% with highest moisture content in puti and lowest in chapila. At the end of the 60 days the moisture content reached to the range of 18.5 to 19.0% which was more or less near the recommended limit of 16% for dried fishery products. The moisture content beyond the recommended limit as the storage period increased further and at the end of 90 days the moisture content increased to the range of 22.9 to 24% when organoleptically the product quality became very poor. The changes in the value of total volatile base nitrogen (TVB-N), peroxide value (PO), moisture and aerobic plate count (APC) of solar tunnel dried products in sealed polythene packages were investigated during 60 days of storage. There was little or no differences in TVB-N, PO and bacterial load of each species packed under various polythene density. The initial TVB-N values were in the range of 10.30 to 12.40 mg/100g of the samples. TVB-N value increased slowly up to the end of the storage period and was to in the range of 46.20 to 57.00 mg/1 00 g of sample. Initially the peroxide values (P.O.) were in the range of 6.54 to 8.40 m.eq./kg oil of the samples. During 60 days of storage, P.O. values increased slowly and at the end of the storage period these values reached to the range of 22.00 to 25.30meq./kg of sample. The initial APC was in the range 5.3xl04-7.3x104 CFU/g. The bacterial load increased slowly and at the end of the 60 days storage period reached to the range 6.6x106 - 8.6x107 CFT/g.

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Fish kamaboko with spinach was prepared by mixing fish kamaboko with spinach prepared with different combinations of ordinary starch (OS) and modified starch (MS), viz., 40:60, 50:50, 60:40, 100%) MS and 100% OS. The products were frozen at -40°C and stored at -20°C, and subjected to biochemical (peroxide value, total titratable acidity, pH and moisture) organoleptic (appearance, taste and colour), microbiological (total plate count) and physical (folding and expressible water) tests at monthly intervals. Among the different combinations tried, it was observed that fish kamaboko with spinach prepared with 50:50 and 60:40 combinations of OS and MS could be stored for six months at-20°C.

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The shelf-life of standardized horse mackerel fish balls was assessed by biochemical, microbiological, organoleptic and other spoilage changes at 0-2°C. There was decrease in pH value, moisture and the organoleptic scores. Expressible water percentage, TMA-N, TVB-N and peroxide value showed increasing trends. Total plate count also increased gradually during storage. Water separation in the treated sample was observed after 12 days and slimy consistency was noticed in the control sample on the 24th day. Based on these observations, it can be concluded that fish balls can be stored at 0-2°C for 20 days.

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The study was conducted on the present status of HACCP based quality management system of golda, Macrobrachium rosenbergii farms in Fulpur region of Mymensingh. Information was collected on general condition of farms, culture systems and post-harvest quality management. In almost all farms, there is no or inadequate infrastructure facilities such as, road access, electric supply, telecommunications, ice, feed storage facility, vehicle for golda transportation, washing and toilet facilities. The problems associated with sanitation and hygiene was: widespread use of cow dung, poultry manure and construction of open toilet within the vicinity of prawn culture pond. Different grades of commercially available and locally prepared feeds were used for golda culture in the pond. Golda post-larvae (PL) of 40-50 days old were stocked with carp species. The price of golda PL ranged from Tk. 1.00 to Tk. 1.25/piece. The pond size varied from 50 decimal (0.2 ha) to 2.5 acre (1.0 ha) with an average depth of 2-2.5 m. The culture period of golda varied from April-May to November-December and survival rate ranged between 75 and 80%. Production of golda varied from 250-500 kg/acre (625-1,250 kg/ha). Harvested golda were transported to city market within 4 h. Two size grading were generally followed during pricing, e.g. Tk. 500 to 550/kg for >100 g size and Tk. 300/kg for <100 g size. The cost-benefit ratio was found to remain around 1:1.25 depending on availability of PL. Water quality parameters such as, water temperature, pH, dissolved oxygen, total alkalinity and chlorophyll a in five golda farms in Fulpur region were monitored. Water temperature ranged from 29°C to 33°C, dissolved oxygen from 2.28 to 4.13 mg/l, pH between 6.65 and 7.94, alkalinity from 44 to 70 mg/l and chlorophyll a concentration from 61.88 to 102.34 µg/l in the five investigated ponds. The Aerobic Plate Count (APC) of the water sample was within the range of 2.0x10^6 - 2.96x10^7 CFU/ml and of soil samples within the range of 6.9x10^6 - 7.73x10^6 CFU/g. Streptococcus sp., Bacillus sp., Escherichia coli, Staphylococcus sp., Pseudomonas sp. and Salmonella sp. were isolated from pond water and sediment. Different feed samples used for golda was analyzed for proximate composition. Moisture content ranged around 14.14-21.22%, crude protein 20.55-44.1%, lipid 4.67-12.54% and ash 9.7-27.69%. The TVB-N values and peroxide values of feeds used as starter, grower and fish meal were found within the acceptable ranges and samples were free from pathogenic organisms. A training was organized for the golda farmers on HACCP, water quality and post-harvest quality management of prawn.

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Aspartate aminotransferase (E.C. 2.6.1.1.) from the skeletal muscle of fresh water fish Cirrhina mrigala has been purified 40 fold by ammonium sulphate fractionation, adsorption on alumina Csub(8) gel and chromatography using DEAE-cellulose column and the properties of the purified enzyme studied. The pH optimum of the enzyme is 7.8. The Km value of aspartic acid and 2-oxoglutaric acid are found to be 2.8 x 10sub(-3) M and 1.0 x 10sub(-4) M respectively. The activity of enzyme is inhibited by p-chloromercurybenzoate, hydroxylamine hydrochloride and sodium cyanide. The inhibition by pchloromercurybenzoate is reversed by reduced glutathione, B-mercaptoethanol and cysteine. Dicarboxylic acids such as maleic acid, malic acid and succinic acid inhibit the enzyme activity. The enzyme is not activated by any of the metal ions tested and heavy metal ions such as mercury and silver strongly inhibit the enzyme activity.

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The influence of different pre-freezing ice storage periods on the biochemical and organoleptic qualities of Indian oil sardines (Sardinella longiceps) in the individual quick frozen (IQF) and block frozen (BF) forms and frozen storage at temperatures of -12°C and -23°C was studied. The shelf-life of the sardines varied between 24 and 2 weeks for samples iced for 0 to 5 days prior to freezing. The deterioration in quality was accompanied by considerable increase in the peroxide value (PV) and free fatty acid (FFA) content and decrease in salt extractability of the proteins. These changes were more rapid at -12°C than at -23°C. BF sardines appeared to be better than IQF samples with respect to the biochemical changes although the differences in overall organoleptic quality were not significant.

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The frozen storage characteristics of fish fingers made out of two different species, differing in lipid content for a period of six months are outlined. The study reveals that the lipid content of the fish meat used for making fish fingers influences the storage pattern in terms of the chemical parameters like peroxide value, thiobarbituric acid value and free fatty acids. The introduction of monosodium glutamate has improved the flavour of the fish fingers. Further, the application of batter on the fish fingers imparted some protective effect in the case of semi-fatty fish.

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Ice-storage study of blood clam (Anadara granosa) meat in direct contact and out of contact (in 200 gauge polyethylene bag) with ice was taken up to assess the amenability of the meat to icing. Changes in moisture, total protein, non-protein nitrogen, α amino nitrogen, total volatile base nitrogen, glycogen, free fatty acid, peroxide value, total bacterial count and coliform count were followed every day. The raw and cooked meat were also subjected to organoleptic evaluation. The study showed that the clam meat can be ice-stored in very good condition out of contact with ice in polyethylene packets for 4 days and in direct contact with ice for 2 days.

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The study showed that less initial moisture with high salt content will be the best condition for enhanced storage life of dehydrated salt mince. Between sample I (10% salt per meat weight) and sample II (15% salt per meat weight) the latter was comparatively better in colour, odour and longer shelf-life. At room temperature the dehydrated salt mince has not showed any increase in total bacterial count. It is also found that the storage life of the salt mince can be enhanced to a significant extent by lowering the moisture content to below 10% and increasing the salt content to above 30%. Peroxide value, free fatty acid value, total volatile nitrogen and trimethylamine registered gradual increase during storage at room temperature for all the three samples. Among the three samples, the sample treated with 0.1% citric acid and 0.125% butylated hydroxy anisole was comparatively better in appearance and showed less rancidity as indicated by TBA values, up to a period of 15 weeks and thereafter all the three samples were almost similar in storage characteristics. Hence, the treatment with citric acid and B.H.A. has apparently not much significance in improving shelf-life and quality of salted dehydrated fish mince.

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The ice-storage characteristics of Catla catla and Labeo fimbriatus are reported. Muscle pH, moisture, total volatile nitrogen, alpha amino nitrogen and peroxide value and also the changes in total bacterial count are studied. C. catla and L. fimbriatus both could be stored in ice for 18 days.