卵黄抗体三种提取方法的比较研究

运用硫酸铵法、聚乙二醇(PEG)法、冷乙醇法对卵黄抗体进行提取,并使用间接竞争酶联免疫吸附试验(ELISA)方法测定所得卵黄抗体的特异性,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)测其纯度,用Nanodrop2000测其蛋白浓度并计算提取量,比较三种方法所得抗体的特异性、纯度和提取量。结果显示,三种提取方法所得卵黄抗体特异性高低依次为硫酸铵法、冷乙醇法、PEG法;纯度高低依次为硫酸铵法、冷乙醇法、PEG法;三种方法的提取量高低依次为PEG法、硫酸铵法、冷乙醇法。经综合分析与评价,硫酸铵法的抗体特异性较好、纯度较高、提取量也较高,且其操作方法相对简便,是比较具有优势的提取方法。     

不同方法提取卵黄抗体效果的比较

为了优化卵黄抗体提取工艺以及产业化生产提供技术参考,从众多方法中初步筛选了较为适宜的水稀释法、辛酸法、乙酸-乙酸钠法、酚沉淀法和海藻酸钠法,并比较了5种方法的提取工艺、生产成本、卵黄抗体提取量及其效价。试验结果表明,辛酸法的提取液体回收率最高,达到171％;辛酸法提取蛋白的浓度最高,为6．6mg／mL;辛酸法和海藻酸钠法提取卵黄抗体的凝集价最高,为1：128。综合各种因素考虑,辛酸法是提取卵黄抗体较为理想的方法。     

牦牛乳中脂肪测定方法比较

用“哥特里一罗紫法”、“酸水解法”和“快速测定法”（乳成份分析仪法）三种方法进行牦牛乳中脂肪含量的测定,经对测定结果的比较分析得出：在测定牦牛乳中的脂肪含量时“哥特里．罗紫法”优于“酸水解法”;而在现场牦牛乳中脂肪含量测定时宜用“乳成分分析仪”进行快速测定。     

利用数码相机和Photoshop 软件非破坏性测定芒果叶面积的简便方法

建立了一种简单、准确、快捷测量芒果叶片面积的新方法。其原理是利用数码相机获取芒果叶片的数字图像,利用Photoshop 图像处理软件计算叶面积,并与传统的叶面积仪测定法、系数法、方格法、鲜重法进行t检验,结果差异不显著。Photoshop 图像法是芒果叶面积测定的一种最佳的准确无损的测量方法。     

改进的表面张力辅助去核法对小鼠卵母细胞去核的研究

为了探讨改进的小鼠表面张力辅助去核法（STA）的可靠性，本试验用改进的表面张力辅助去核法与表面张力辅助去核法对小鼠卵母细胞进行去核操作，比较其去核操作效率。结果发现，改进的表面张力辅助去核法能显著减少去核操作时间，去核率与表面张力去核法无显著差异。改进的小鼠表面张力辅助去核法是一种适用于小鼠卵母细胞操作的快速、简单、实用的去核方法。     

不同采集方法对猪卵母细胞的采集效率和成熟率的影响

本研究旨在开发出一种对COCs损伤小且采集效率和卵母细胞成熟率高的卵母细胞采集方法.从屠宰场采集来的卵巢运回实验室后随机分为3组,分别用抽吸法、解剖法和刀切过滤法3种方法采集COCs,然后进行体外成熟培养,通过对COCs采集效率、COCs完整性及卵母细胞体外成熟率的比较,研究解剖法、刀切过滤法和抽吸法3种采集方法对猪卵母细胞的采集效率和成熟率的影响.结果表明:每个卵巢采集COCs个数,刀切过滤法显著高于解剖法和抽吸法(P＜0.05),而解剖法仅为1.55个,显著低于抽吸法(P＜0.05);采集效率抽吸法最佳,采集和挑选单个COCs的时间显著低于解剖法和刀切过滤法(P＜0.05),刀切过滤法次之,但显著低于解剖法(P＜0.05);A、B级COCs比例,解剖法高达99.23%,显著高于其他2种方法(P＜0.05),刀切过滤法与抽吸法差异不显著(P＞0.05);卵母细胞的成熟率,解剖法和刀切过滤法都达到80%,二者间差异不显著(P＞0.05),但显著高于抽吸法(P＜0.05);100个成熟卵母细胞所需的卵巢数量,刀切过滤法最少,仅为17.4个,而解剖法和抽吸法分别高达80.6和90.3个;需要的总采集时间(采集和挑选的时间),刀切过滤法最低,仅为126.6 min,而解剖法最高,为758.9min.综上所述,与抽吸法和解剖法相比,刀切过滤法是一种高效、快速的卵母细胞采集方法.     

鸡球虫病活疫苗不同卵囊计数方法的比较

为了解鸡球虫活疫苗不同卵囊计数方法对检测结果的影响,分别采用血细胞计数板4角方格法和9方格法以及麦氏计数板法对3种鸡球虫活疫苗各进行了6次卵囊计数,然后采用t检验和Z比分值方法比较筛选出最适宜的计数方法,并对检测方法进行了验证和变异系数统计。结果显示：采用每批检品抽检1瓶,通过血细胞计数板4角方格法重复计数6次,去除最大值和最小值后取平均值作为该瓶检品卵囊计数结果,如果首次检验结果不符合规定可重检一次的方法,人员误差最小。该方法提高了鸡球虫活疫苗卵囊计数结果的可重复性和准确性,增强了疫苗质量控制的可靠性。     

银狐垂体细胞系构建研究

分别采用组织块法和消化法2种接种方法进行银狐垂体细胞系构建研究。在消化法中使用不同浓度的胰蛋白酶来检验效果,同时在原代培养期使用反复差速贴壁法纯化,传代培养期结合使用反复差速贴壁以及两步消化法进行处理。结果表明：组织块法接种后成纤维细胞生长明显,原代培养后期成纤维细胞已为主要细胞;降低胰蛋白酶浓度能降低对细胞损伤,但消化时间会增长;采用消化法接种在抑制成纤维细胞生长方面效果好于组织块法。纯化效果上,结合反复差速贴壁法与两步消化法处理后获得了较好的效果,在原代培养时期能较好去除成纤维细胞并通过持续使用该方法可在传二代后获得较高纯度的细胞,纯度能达到80%以上,能够建立细胞系。     

微生物浊度法与管碟法测定吉他霉素效价的比较研究

为比较微生物浊度法和管碟法测定吉他霉素效价,以《中国药典》(2015年版)吉他霉素抗生素微生物检定法浊度法和《中国兽药典》(2010年版)吉他霉素抗生素微生物检定法管碟法为依据测定一批吉他霉素效价。结果显示:浊度法测定效价平均值为1691 U/mg,RSD为0.9%;管碟法测定效价平均值为1687 U/mg,RSD为1.0%,两种方法测定结果无显著性差异(P0.05)。本实验表明,浊度法测定吉他霉素效价与管碟法相比更加快速、便捷,而且可避免吉他霉素多组分的影响因素。     

Modified flotation method with the use of Percoll for the detection of Isospora suis oocysts in suckling piglet faeces

The modification of flotation method for the examination of diarrhoeic piglet faeces for the detection of Isospora suis oocysts was elaborated. The method was based on removing fractions of fat from the sample of faeces by centrifugation with a 25% Percoll solution. The investigations were carried out in comparison to the McMaster method. From five variants of the Percoll flotation method, the best results were obtained when 2ml of flotation liquid per 1g of faeces were used. The limit of detection in the Percoll flotation method was 160 oocysts per 1g, and was better than with the McMaster method. The efficacy of the modified method was confirmed by results obtained in the examination of the I. suis infected piglets. From all faecal samples, positive samples in the Percoll flotation method were double the results than that of the routine method. Oocysts were first detected by the Percoll flotation method on day 4 post-invasion, i.e. one-day earlier than with the McMaster method. During the experiment (except for 3 days), the extensity of I. suis invasion in the litter examined by the Percoll flotation method was higher than that with the McMaster method. The obtained results show that the modified flotation method with the use of Percoll could be applied in the diagnostics of suckling piglet isosporosis.     

基于数码相机和Auto CAD软件测定园林植物叶面积的简便方法

提出了一个快速的非破坏性叶面积测定方法,即采用数码相机获取叶片的数字图像,用AutoCAD软件计算园林植物叶面积。并与目前常用的网格法、剪纸法和叶面积仪测定法进行比较分析。研究结果表明,该方法特别适合于针叶、小叶类园林植物以及草坪草叶面积的测定。     

Precision of estimated QTL positions in granddaughter designs using combined haplotype sharing TDT and linkage analysis

The aim of this study was to develop the linear haplotype sharing transmission disequilibrium test (LHS-TDT) method and combine this method with the simple regression method to estimate the precision of QTL positions in granddaughter designs. This precision was determined by Monte Carlo simulation in granddaughter designs. A single bi-allelic QTL at the midpoint of a linkage group and 26 markers with 1 cM intervals and with two alleles each were simulated. Three linear models, (i.e. the simple regression model, the linear haplotype sharing TDT method and the combination of these two models) were compared. The mean of absolute differences (A) between the estimated and true QTL position of each method was considered for six different scenarios consisting of combinations of a number of markers and the most frequent haplotypes. The mean of A, using the simple regression method, was 4.38 centimorgan (cM). The means of A using the LHS-TDT method were less than the simple regression method in all scenarios and ranged from 1.86 to 3.82 cM depending on the scenario. The mean of A using the combined method was more than the LHS-TDT method and less than the simple regression method. The means of A using the combined method ranged from 2.32 to 4.36 cM. Therefore, for populations similar to those population simulated in this study, the LHS-TDT was better than the simple regression method and the combined method for precision of estimated QTL position in granddaughter designs.     

压力厚度法测量獭兔被毛密度

分别采用直接计数法、毛囊组织切片法和压力厚度法3种方法对4组獭兔被毛密度进行测定,通过综合比较分析测定结果对测定方法进行评估。结果表明:直接计数法耗时多,取样难度大。组织切片法相对复杂,需要专用设备。这两种方法对獭兔毛皮都会造成一定损伤。压力厚度法能快速测量被毛密度,其效率较高,且不会破坏样品。     

鹅细小病毒PCR检测方法的建立及应用

根据鹅细小病毒VP3基因设计合成一对引物,结果特异性地扩增出与预期片段大小相符的430bp核苷酸片段,建立了用于检测鹅细小病毒的PCR方法。此方法检测鹅细小病毒结果与病毒分离培养、电镜检查结果一致。通过特异性、敏感性及临床应用实验证明,每5μL样品中获得阳性扩增结果中最小检出量为2．5×10^-1.58 ELD50;此方法用于临床检测鹅细小病毒是特异的、敏感的、可行的,本次实验建立的PCR方法使我省小鹅瘟的诊断,监测和防制工作上升到分子生物学水平。     

鹌鹑肠道微生物总DNA的最佳提取方法

为了获取高质量、较完整的肠道菌群基因组DNA,试验采用常规的饱和苯酚/氯仿法(方法一)、牛瘤胃DNA的提取法(方法二)和对上述两种方法进行优化后得到的肠道微生物DNA的提取法(方法三)对鹌鹑肠道微生物总DNA进行了提取,并对结果进行了比较。结果表明:应用方法一和方法二提取的DNA浓度比较低,电泳显示样品DNA条带没有方法三明亮;以方法三提取的肠道总DNA为模板,采用细菌通用引物,对其16S rDNA V3可变区进行PCR扩增,得到较清晰的图谱,条带整齐。说明采用方法三提取鹌鹑肠道微生物DNA较完整,可用于后续的分子生物学试验研究。     

2种鸭蛋中胆固醇测定方法的比较

为确定蛋黄样品中的蛋白质对鸭蛋中胆固醇测定结果的影响程度,采用直接试剂盒法和间接试剂盒法对市售鸭蛋中的胆固醇含量进行了测定,并对2种方法的测定结果进行了比较。结果表明,直接法和间接法的测定值具有极显著的相关性,但直接法测定值较间接法测定值高。     

试论地方猪品种的现代保种方法

本文讨论了以群体遗传学理论为基础的现行地方猪品种的保种方法，指出了该方法的不足之处，同时也分析了以分子遗传学为基础的分子遗传标记辅助保种法的可行性，说明该方法可以弥补现行地方猪品种保种方法的不足之处，指出现代地方猪品种的保种方法应该是二者的结合。     

Variational bayesian method of estimating variance components

We developed a Bayesian analysis approach by using a variational inference method, a so‐called variational Bayesian method, to determine the posterior distributions of variance components. This variational Bayesian method and an alternative Bayesian method using Gibbs sampling were compared in estimating genetic and residual variance components from both simulated data and publically available real pig data. In the simulated data set, we observed strong bias toward overestimation of genetic variance for the variational Bayesian method in the case of low heritability and low population size, and less bias was detected with larger population sizes in both methods examined. The differences in the estimates of variance components between the variational Bayesian and the Gibbs sampling were not found in the real pig data. However, the posterior distributions of the variance components obtained with the variational Bayesian method had shorter tails than those obtained with the Gibbs sampling. Consequently, the posterior standard deviations of the genetic and residual variances of the variational Bayesian method were lower than those of the method using Gibbs sampling. The computing time required was much shorter with the variational Bayesian method than with the method using Gibbs sampling.     

食草动物食性研究的主要方法及其评价

本文对国内外常用的食草动物食性研究方法进行了分析与比较。常见的研究方法主要包括胃分析法、室内饲喂法、粪便显微组织学分析法、直接观察法和利用法。粪便显微分析法分析结果较为精确,并且易于取样,相比较更适用于我国食草动物的食性研究。胃分析法要求研究者必须杀死动物,利用法精确性较差,而观察法多受研究条件的限制, 因此这3种方法多用于食物组成的定性描述,或是作为其它研究方法的补充。     

Counting coccidial oocysts in chicken faeces: a comparative study of a standard McMaster technique and a new rapid method

For the assessment of coccidial oocyst production by chickens, some modified form of the McMaster counting method is commonly used. The objective of this study was to evaluate a standard method and to compare it to a new, faster method, in which all the preparative stages before counting are carried out in the same container into which the original faecal sample was collected. A stock suspension containing purified oocysts of all seven valid Eimeria species that parasitize chickens was prepared, from which seven concentrations of oocyst suspensions were made. Since the faecal material in a sample influences the ability of oocysts to float up in a McMaster chamber, the new method was tested to establish the optimal amount of faeces in the original sample. Control oocyst suspensions containing no faeces were also tested, and three series of counts using the new method were compared with the standard McMaster method. The results were statistically analysed by agreement analysis. Repeatability and between-operator variation of both methods were also tested by agreement analysis. Counting by the standard McMaster method underestimated the true number of oocysts. The new method gave counts in agreement with the true number of oocysts if using 1 g of faeces per sample. With 2 g of faeces, counts were obtained that agreed with counts by the standard McMaster method. Both methods showed agreement between repeated measurements. The new method used on a sample containing 2 g of faeces provides a convenient alternative to the standard modified McMaster method. A 1-g faecal sample increases agreement with the true numbers of oocysts. Processing of a sample with the new method is about nine times faster than with the standard method.