共查询到19条相似文献，搜索用时 203 毫秒
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酸性α-醋酸萘酯酶染色法在鳜细胞免疫水平检测中的应用研究 总被引：1，自引：0，他引：1
研究了酸性α-醋酸萘酯酶染色法在鳜细胞免疫水平检测中应用的可行性。用不同比重的淋巴细胞分离液对鳜外周血中的淋巴细胞进行分离,结果表明比重为1.080的分离液可最大程度地将鳜外周血中的淋巴细胞分离出来。用酸性α-醋酸萘酯酶染色法对分离出的淋巴细胞进行染色,结果显示最佳的染色条件为在pH6.0~6.5的孵育液中37℃孵育90 m in,然后2%甲基绿37℃复染10 m in,酸性α-醋酸萘酯酶阳性细胞胞核被染成绿色而胞浆被染成红色,阴性细胞胞核和胞浆都被绿染。对正常未免疫鳜与嗜水气单胞菌脂多糖和鳜病毒主要衣壳蛋白免疫一周后的鳜外周血淋巴细胞染色计数,结果酸性α-醋酸萘酯酶阳性细胞所占的比例分别为15%~18%、33%~40%和26%~29%,方差分析表明免疫组与对照组阳性率差异极显著,说明α-醋酸萘酯酶染色法可用于鳜细胞免疫水平的检测。 相似文献
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采用紫外分光光度法测定栉孔扇贝（Chlamysfarreri）血淋巴中酯酶的活性，并研究底物、抑制剂、激活剂、温度、pH对酯酶活力的影响及酶的热稳定性。结果表明，栉孔扇贝血清与血细胞中均存在酯酶，且血细胞中的酶活力高于血清。血清与血细胞中酯酶的最适作用底物有所不同，血清中为α-醋酸萘酯，而血细胞中为α-丁酸萘酯。当以α-醋酸萘酯为底物时，血清中酯酶的最适温度为45℃，最适pH值为7．5；以α-丁酸萘酯为底物时，血清中酯酶的最适温度为40℃，最适pH值为8．0。以α-醋酸萘酯及α-丁酸萘酯为底物分别测定酶的热稳定时，酶活力变化相似，在20℃时活力最高，而后缓慢下降。栉孔扇贝血淋巴中酯酶属中温型中性酶类，低温下较稳定，底物专一性不强，血清中的酯酶可能来自血细胞，该酶在贝类的免疫防御中町发挥重要的作用。 相似文献
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《畜禽业》2001,(8):55
佛山科学技术学院冯军等选择1日龄石歧杂黄鸡150只,健康观察3d,第4d按体重随机分为3组.试验Ⅰ、Ⅱ组从4～42d分别饲喂含氟量300mg/kg和500mg/kg的饲料,43～56d喂常规饲料,对照组整个试验期内均喂常规饲料.于14d、28d、42d、56d时,用酸性α-醋酸萘酯酶(ANAE)和形态学检查法测定外周血T-淋巴细胞(T-LT)ANAE阳性率和淋巴细胞(LT)转化率,以反映鸡的细胞免疫状况.结果表明:在4～8周内,试验组鸡只增重减慢,胸腺器官指数降低,外周血淋巴细胞ANAE阳性率和淋巴细胞转化率显著下降,表明氟化物能对细胞免疫功能呈现明显的抑制作用,该抑制作用可能是由于胸腺生长发育受阻,从而影响了淋巴细胞的诱导、增殖、分化.
<畜牧与兽医>,2001(2):10～12 相似文献
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分别采用海藻酸钠包埋法、明胶包埋交联法、壳聚糖吸附交联法制备固定化海洋芽孢杆菌酯酶ETM-b,并对其固定化条件进行了研究。结果发现,壳聚糖制备的固定化酶效果最好,壳聚糖2%、戊二醛浓度1%、小球与酶液4∶3(g/ml)时制备的固定化酶的活性回收最高,达到66%。壳聚糖制备的固定化酶使用10次,相对活性保留70%,具有良好的操作稳定性。固定化酶在非水介质中具有转化α-乙酸萘酯(α-Naphthyl acetate)的能力,在异辛烷、正辛烷、正己烷中活性表现最高。 相似文献
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《福建水产》2020,(2)
研究建立了基于顶空气相色谱-质谱联用(HS-GC/MS)的裂解碳九主要成分分析方法,该方法可同时测定水体中α-甲基苯乙烯、间-甲基苯乙烯、对-甲基苯乙烯、邻-甲基苯乙烯、β-甲基苯乙烯、双环戊二烯、茚、1-甲基萘和2-甲基萘等9种裂解碳九主要成分的残留量。取10.0 mL海水样品在密封的顶空瓶中,在振荡器中于90℃下振荡10 min,使气-液两相达到动态平衡。采用DB-5MS毛细管色谱柱(30 m×0.25 mm×0.25μm)对9种目标化合物进行分离,内标法定量。本方法在0.50～10μg·L~(-1)范围内线性关系良好,相关系数均大于0.998;在海水中9种化合物的检出限为0.500～1.00μg·L~(-1),添加浓度为1.00～10.0μg·L~(-1)时,各目标化合物的平均加标回收率在82.4%～106.6%之间,相对标准偏差(n=6)在3.73%～9.41%之间。本方法操作简单、快速,灵敏度和准确度高,适用于海水中裂解碳九主要成分残留量的同时检测,为海洋污染事故应急处置分析中裂解碳九主要组成的测定提供参考依据。 相似文献
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海虾超高压灭菌的试验研究 总被引：1，自引：1，他引：1
以新鲜海虾(Fenneropenaeus chinensis)为对象,研究了重复加压对超高压灭菌效果的影响.采用正交试验设计方法考察了不同工艺参数(处理压力、保压时间、加压次数)对海虾中微生物存活量的影响,确定超高压处理海虾的最佳工艺条件.试验结果表明,当处理压力为300～400 Mpa,保压时间为10～20 min,加压2~3次时,对海虾中各种微生物杀灭作用显著.在重复3次加压,压力为400 Mpa,保压时间为15 min的条件下,可以杀灭海虾中99.3%的微生物. 相似文献
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通过以三丁酸甘油酯和α-乙酸萘酯为底物,建立了一种快速、准确的酯酶产生菌的筛选方法,并从样品中分离到 1 株产酯酶活力和稳定性较高的菌株 EB-1,对其进行了形态学、生理生化特征鉴定和 16S rDNA 序列分析.结果表明,该菌株与Bacillus subtilis subsp. subtilis 模式菌株的同源性高达 99.79%;又根据其产黑色素这一特性,最终将其确定为Bacillus subtilis var. niger,其 16S rDNA 序列在 GenBank 的注册号为 EU016526. 相似文献
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为减少甲壳素生产过程中废酸液对环境的污染,降低甲壳素的生产成本,试验以甲壳素生产过程中的废盐酸为研究对象,利用硫酸和氯化钙反应进行回收、循环利用盐酸溶液,通过单因素试验和正交试验对工艺条件进行优化。结果表明,脱钙废盐酸溶液最佳再生条件:反应温度为室温,硫酸质量分数为98%,硫酸与废盐酸中钙离子的摩尔比为2.3∶1,反应时间13 min,静置时间3.5 h,最佳条件下盐酸再生率为91.51%。利用最佳工艺条件对废盐酸进行再生处理,盐酸可循环重复利用5次,脱钙虾壳的碳酸钙残留量≤1.2%。 相似文献
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Carolina Fernandez Dario Mascolo Sean J. Monaghan Johanna L. Baily Lynn Chalmers Giuseppe Paladini Alexandra Adams James E. Bron Sophie Fridman 《Journal of fish diseases》2019,42(6):883-894
Two aqueous fixation methods (modified Davidson's solution and modified Davidson's solution with 2% (w/v) Alcian blue) were compared against two non‐aqueous fixation methods (methacarn solution and methacarn solution with 2% (w/v) Alcian blue) along with the standard buffered formalin fixation method to (a) improve preservation of the mucous coat on Atlantic salmon, Salmo salar L., gills and (b) to examine the interaction between the amoebae and mucus on the gill during an infection with amoebic gill disease. Aqueous fixatives demonstrated excellent cytological preservation but failed to deliver the preservation of the mucus when compared to the non‐aqueous‐based fixatives; qualitative and semi‐quantitative analysis revealed a greater preservation of the gill mucus using the non‐aqueous methacarn solution. A combination of this fixation method and an Alcian blue/Periodic acid–Schiff staining was tested in gills of Atlantic salmon infected with amoebic gill disease; lectin labelling was also used to confirm the mucus preservation in the methacarn‐fixed tissue. Amoebae were observed closely associated with the mucus demonstrating that the techniques employed for preservation of the mucous coat can indeed avoid the loss of potential mucus‐embedded parasites, thus providing a better understanding of the relationship between the mucus and parasite. 相似文献
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R. Eckmann 《Fisheries Management and Ecology》2003,10(4):233-239
Alizarin red S from 200 to 5000 mg L?1 was used to mark the otoliths of whitefish embryos Coregonus lavaretus (L.) from 5 days after fertilization until shortly before hatching, to develop a method for evaluating the effect of stocking whitefish. Mark quality depended on developmental stage at the time of immersion. The best results were obtained when the otolith primordia had formed and had started aggregating to form the innermost part of the nucleus. This was verified by in vitro staining of otoliths where the primordia were more intensely stained than the remaining part of the nucleus. The volume ratio eggs to ARS solution, which was 1:10 in all but two treatments, must be observed when comparing fluorochrome concentrations, as mark quality decreased at higher ratios of 1:5 and 1:2. An average mortality rate of 35% in the most successful treatments was considered tolerable for a mass‐marking programme being used under commercial hatchery conditions. This method for marking otoliths of whitefish embryos with ARS during egg incubation is recommended for all cases where marking of larvae is not possible. 相似文献
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Joshua Woolsey Michael Holcomb Joseph G Cloud & Rolf L Ingermann 《Aquaculture Research》2006,37(3):215-223
This study examined the pH sensitivity of steelhead, Oncorhynchus mykiss (Walbaum), sperm motility relative to the composition of incubation and activation media. The percentage of sperm that initiated motility following incubation in a sperm immobilizing solution (SI) titrated to different pH values and subsequent activation by dilution in buffered swimming medium (SM) at pH 8.5 or 50% ovarian fluid (OF) showed little or no pH sensitivity; sperm diluted in de‐ionized water (DI) showed no motility after incubation at any pH. In contrast, motility of sperm diluted in tap water (TAP) was highly sensitive to the pH of the incubation medium. Sperm incubated with buffered seminal plasma at high, but not low pH demonstrated high percent motility when diluted with DI. Sperm incubated in low‐pH SI demonstrated high motility only when diluted into high‐pH SM. The effects of the composition of incubation and activation media on sperm motility were generally reflected in comparable effects on fertility. Therefore, these data indicate that the pH sensitivity of sperm motility and fertility depends on the composition of commonly used incubation as well as activation media. 相似文献
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为解决当前低黏度及超低黏度褐藻胶生产工艺中存在的不足,本实验以海带为原料,利用褐藻胶裂解酶降解制备低黏度及超低黏度褐藻胶,研究了分子量、p H、温度对黏度的影响,确定了碱消化的最佳条件,探究了酶解工艺中加酶量、酶解时间及原料的初始黏度对褐藻胶产品的影响。结果显示,通过控制加酶量(100~500 U/g),酶解30 min即可得到低黏度及超低黏度褐藻胶,其中加酶量为100~330 U/g时可得到低黏度褐藻胶,加酶量增加至330~500 U/g时,可得到超低黏度褐藻胶,且酶解法得到的褐藻胶样品分子量均一度高,工艺节水率高达10%~50%;同时研究发现酶解样品黏度与原料初始黏度相关性不大,只在较短时间内表现出相关性,该工艺具有较高的原料适用性。 相似文献
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尼罗罗非鱼卵黄脂磷蛋白的分离纯化与性质鉴定 总被引：1，自引：1，他引：0
采用Sephacryl S-300过滤层析和DEAE-Sepharose Fast Flow离子交换层析相结合的方法从尼罗罗非鱼(Oreochromis niloticus)成熟卵子匀浆液中分离纯化出了一种高分子量的蛋白。该蛋白能被Schiff试剂、甲基绿和苏丹黑B着色,Western blot显示能被金鱼卵黄脂磷蛋白(lipovitellin,Lv)多克隆抗血清特异性识别,在非变性条件下分子量约为560 k D,在SDS变性条件下分子量约为112 k D,结果表明分离纯化的蛋白是一种含有糖、磷、脂基团的蛋白,符合鱼类Lv的性质,且与金鱼Lv有免疫交叉反应,从蛋白的性质和免疫原性以及分子量大小等角度判断,本研究获得的高纯度蛋白为尼罗罗非鱼卵黄脂磷蛋白;纯化的罗非鱼Lv在反复冻融、37℃及60℃处理条件下均未出现降解,表明罗非鱼Lv比鱼类卵黄原蛋白(Vitellogenin,Vtg)更为稳定。研究结果为罗非鱼Lv抗体的制备奠定了基础。 相似文献
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Extreme ocean acidification reduces the susceptibility of eastern oyster shells to a polydorid parasite 
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下载免费PDF全文 J C Clements D Bourque J McLaughlin M Stephenson L A Comeau 《Journal of fish diseases》2017,40(11):1573-1585
Ocean acidification poses a threat to marine organisms. While the physiological and behavioural effects of ocean acidification have received much attention, the effects of acidification on the susceptibility of farmed shellfish to parasitic infections are poorly understood. Here we describe the effects of moderate (pH 7.5) and extreme (pH 7.0) ocean acidification on the susceptibility of Crassostrea virginica shells to infection by a parasitic polydorid, Polydora websteri. Under laboratory conditions, shells were exposed to three pH treatments (7.0, 7.5 and 8.0) for 3‐ and 5‐week periods. Treated shells were subsequently transferred to an oyster aquaculture site (which had recently reported an outbreak of P. websteri) for 50 days to test for effects of pH and exposure time on P. websteri recruitment to oyster shells. Results indicated that pH and exposure time did not affect the length, width or weight of the shells. Interestingly, P. websteri counts were significantly lower under extreme (pH 7.0; ~50% reduction), but not moderate (pH 7.5; ~20% reduction) acidification levels; exposure time had no effect. This study suggests that extreme levels – but not current and projected near‐future levels – of acidification (?pH ~1 unit) can reduce the susceptibility of eastern oyster shells to P. websteri infections. 相似文献
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为建立1种尼罗罗非鱼肌肉组织蛋白质的双向电泳体系,实验将尼罗罗非鱼肌肉组织蛋白质提取后,用双向电泳(2-DE)分离蛋白质,分别对蛋白质样品的制备方法、IPG胶条长度及p H范围、SDS-PAGE凝胶浓度及染色方法4个关键因素进行了探索和优化。结果显示,采用液氮研磨-丙酮沉淀法制备样品蛋白质,使用24 cm p H 4~7的IPG胶条进行第一向等电聚焦电泳,第二向SDS-PAGE电泳采用浓度为12.5%的凝胶进行,双向电泳后的凝胶采用硝酸银染色法进行染色处理,该条件下扫描所得尼罗罗非鱼肌肉蛋白质二维电泳图谱具有蛋白质分离程度好、斑点清晰、分辨率高及横纹少等优点。研究表明,技术体系适用于尼罗罗非鱼肌肉蛋白质的分离,可用于后续罗非鱼肌肉蛋白质组学研究。 相似文献
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Survival of Aeromonas salmonicida was monitored during prolonged incubation in either distilled water or lake water. Culturability was determined from colony forming units enumerated on tryptone soy agar, whilst flow cytometry was used for direct analysis of viable cells after staining with fluorescent dyes which differentially stained bacteria in relation to defined cellular properties. Over time, populations of culturable cells steadily declined and were not detected after 10 days incubation in distilled water or 33 days in lake water. Flow cytometric analysis revealed that cellular properties related to viability were lost shortly after culturable cells became undetectable in distilled water. In contrast, those incubated in lake water showed little change in these properties over a 57-day experimental period. The implications of these differences are discussed, and it is concluded that A. salmonicida is capable of remaining intact and active upon prolonged incubation in lake water, although this does not conclusively prove viability. 相似文献