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1.
用犊牛睾丸细胞培养传染性牛鼻气管炎病毒(IBRV),反复冻融,差速离心提取病毒,用Triton X-100溶解,超声波处理,制成IBRV TritonX-100亚单位抗原,经SDS-PAGE电泳,其分子量在176kD-53kD之间,其中有6条清晰的蛋白带。该抗原与一定比例的白油-司盘佐剂乳化,接种育成年,经间接ELISA检测,可使接种的育成牛产生高效价的血清抗体(OD492mm=1.57)。2种不同剂量(80mg/头,40mg/头)的IBRV亚单位疫苗接种育成牛,体内抗体效价相差不显著,抗体可在体内持续12周,用IBRV TrionX-100亚单位疫苗2次接种育成牛,其体内抗体效价显著高于用灭活疫苗2次接种育成牛体内的效价。试验结果表明:提取的IBRV多肽制作的亚单位疫苗具有良好的免疫原性,且抗体持续时间较长。  相似文献   
2.
本文应用间接法Dot-ELISA进行鸡传染性法氏囊病病毒(IBDV)抗原的检测,其最低检出量为0.43μg/ml。对人工感染/自然病例的各组织脏器进行检测,结果法氏囊的IBDV检出率为100%,脾脏的检出率为80%/84.9%,其它器官组织如肾、胸腺、心、肝、肺、盲肠扁桃体、胸肌等均含有一定量的病毒(11~60%)。经统计学分析,法氏囊与脾脏的病毒检出率之间具有显著性差异(人工感染差异极显著P<0.01,自然病例差异显著P<0.05),进一步证实法氏囊是IBDV最主要的靶器官。试验结果表明,该法检测IBDV抗原,敏感性高、特异性强、重复性好,并且经济、方便、快速,适于大批量样品检测,可作为一种诊断IBD的比较理想的方法。  相似文献   
3.
20世纪末以来,经济和科技两方面的发展使证券交易体制发生了从直接持有体制到间接持有体制的革命性转换。在证券的间接持有体制下,证券投资者与中介机构之间的法律关系是行纪关系;证券投资者对证券享有的权属的性质是“所有权”。由于同一托管人为众多的证券投资者托管,并且证券投资者对证券享有的所有权是通过支配不包含有体物的财产权行使的,所以证券投资者对证券享有的是一种“按份共有”形式的“准所有权”。  相似文献   
4.
The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.  相似文献   
5.
枣树间接地下滴灌根区土壤盐分运移规律研究   总被引:7,自引:0,他引:7  
通过小区试验研究了间接地下滴灌条件下密植枣树根区不同方向(水平及垂直方向)、不同土层盐分质量比的变化特征,及导水装置直径(50、75、90 mm)和灌水量(9、13、17 L/(棵·次))对枣树根区土壤盐分运移的影响。结果表明:经过一段时间后,垂直方向10~40 cm土层盐分质量比较低,0~10 cm土层盐分质量比最高;水平方向10~50 cm各土层盐分质量比随与出水口水平距离的增加而增大。在单次灌水后导水装置直径一定时各土层脱盐率随灌水量的增加而增大。当灌水量一定时,10~40 cm土层脱盐率随导水装置直径的增加呈减小的趋势,0~10 cm、40~50 cm脱盐率随导水装置直径增大呈低-高-低的趋势。在7—9月份,当导水装置直径为75 mm,灌水量为13、17 L/(棵·次)时,0~50 cm土层土壤平均盐分质量比均较低而且变化幅度较小。综合考虑节水、抑盐效果,确定导水装置直径为75 mm,灌水量为13 L/(棵·次)为适宜的间接地下滴灌灌溉模式。  相似文献   
6.
Chlamydia spp. are obligate intracellular gram-negative bacteria that cause a wide range of significant diseases in humans and animals worldwide, resulting in significant economic losses. Chlamydial infection in cattle has been reported in many countries including China. However, there has been no survey of chlamydial infection of dairy cattle in Guangzhou, southern China. The objective of the present investigation was to examine the chlamydial seroprevalence in dairy cattle in Guangzhou, subtropical southern China by using an indirect hemagglutination assay (IHA). The overall seroprevalence of chlamydial infection in dairy cattle was 7.25% (29/400). Greater than or equal to eight-yr-old dairy cattle had the highest seroprevalence (10.34%), followed by those that were ≥ 6 years old or < 7 years old dairy cattle (10.20%), although there were no statistically significant differences among different groups (P > 0.05). Dairy cattle with 5 pregnancies had the highest seroprevalence (10.81%). These results indicate that chlamydial infection was present in dairy cattle in Guangzhou, subtropical southern China, and integrated strategies and measures should be executed to control and prevent chlamydial infection and disease outbreak in the study region.  相似文献   
7.
The effects of a novel pyrimidine herbicide, NS-245852 [2-chloro-6-fluorophenyl-4-(trifluoromethyl)thieno[2,3-d]pyrimidine-2-yl-ketone], on mitosis in oat ( Avena sativa L. cv. Zenshin) root tips were investigated by using light and immunofluorescence microscopy. The root growth was strongly inhibited at 10−7 mol L−1 of NS-245852, and swollen root tips were induced at 5 × 10−8 mol L−1. As observed by the use of light microscopy, the herbicide produced disrupted mitosis and large polynucleate cells in the meristematic root tissue. These symptoms were similar to those of mitotic disrupter herbicides. The immunofluorescence microscopy studies of the root tip cells treated for 30 min revealed that spindle fibers and the preprophase band were reduced, although kinetochore fibers and the phragmoplast were not affected. Kinetochore fibers remained as small fluorescence spots, and the phragmoplast disappeared after a 3 h treatment. No microtubule arrays were observed by a longer treatment (longer than 3 h). Among the microtubule arrays, spindle fibers and the preprophase band were found to be the most sensitive to the herbicide, whereas kinetochore fibers were the most resistant. The phragmoplast was intermediate. Thus, the primary action of NS-245852 is the inhibition of polymerization of tubulin into microtubules.  相似文献   
8.
The use of flow cytometry in veterinary diagnostics is becoming a valuable clinical tool with a broad range of applications. Physical characteristics of cells can be determined by the flow cytometer laser and electronics through the measurement of changes in light scatter properties. Other components and functions of cells can be defined through the application of fluorochrome dyes that have an affinity for cellular components. Traditionally, common clinical applications are immunophenotyping of cells of the hematopoietic system with fluorescent-labeled antibodies raised against specific cell surface proteins. Other approaches have been used to elucidate changes in cell function and DNA content. This review is intended to provide the reader with the fundamental uses of flow cytometry. Examples of clinical applications in equine patients include immune-mediated hemolytic anemia, immune-mediated thrombocytopenia (IMT), chronic inflammatory disease, and neoplasia.  相似文献   
9.
The gammaherpesvirus bovine herpesvirus-4 (BHV-4) has been isolated from a wide variety of animals, including lions and domestic cats. Although BHV-4 antibodies have been detected in normal cats and cats with urinary disorders, the epidemiology and pathogenic role of BHV-4 in cats is unknown. The purpose of this study was to determine the prevalence of BHV-4 antibodies and viral nucleic acid in a population of free-roaming cats. Plasma and peripheral blood leukocyte samples were collected from 52 male and 52 female free-roaming cats impounded at a regional animal control facility in Central Michigan. Plasma concentrations of BHV-4 antibodies were measured with an indirect fluorescent antibody test. Peripheral blood leukocyte DNA was isolated, and a 2-stage polymerase chain reaction with heminested primers delineating a conserved portion of the BHV-4 glycoprotein B gene homologue was used to amplify BHV-4-specific DNA sequences. BHV-4 antibodies were detected in 38 (73%) male and 23 (44%) female cats. Seropositive cats were significantly more likely to be male than female (odds ratio = 3.22; P = .007). Cell-associated viremia was detected in 17 (33%) male and 11 (21%) female cats. Of the 61 seropositive cats, 23 (38%) had a detectable viremia; only 5 (12%) seronegative cats had detectable viremia. Seropositive cats were significantly more likely to be viremic than seronegative cats (OR = 4.30: P = .009). Our results suggest that BHV-4 infection may be more widespread in certain cat populations than previously reported. Furthermore, many cats seropositive for BHV-4 antibodies have a concurrent cell-associated viremia.  相似文献   
10.
Classes of antibody bound to erythrocytes were determined using direct immunofluorescence (DIF) flow cytometry in 3 horses and 12 dogs with immune-mediated hemolytic anemia (IMHA). Background levels of antibody binding were determined in samples from 12 horses and 12 dogs that were free of clinical disease. The range of nonspecific binding of a fluorescein isothiocyanate (FITC)-conjugated goat anti-equine immunoglobulin G (IgG) was 19.9–36.7%, but was eliminated by the use of the F(ab)2 fragment of FITC-conjugated goat anti-equine IgG. Background binding by other class-specific antibodies to equine and canine erythrocytes was negligible. The DIF results were compared to the direct antiglobulin (Coombs) test in 5 horses and 20 dogs with anemia. The former assay was more sensitive in dogs with IMHA than was the Coombs' test (100% versus 58%). In contrast, the Coombs' test had better specificity than the DIF assay (100% versus 87.5%, respectively). Using clinical parameters or response to therapy as the comparison, the positive and negative predictive values for the DIF test were 92% and 100% compared to the values of the Coombs' test of 100% and 62%. The DIF assay detected low levels of cells bound with antibody (<30%) in 5 dogs that were Coombs' test-negative. For both species, performance of the DIF test was independent of the prozone effect. Five dogs with IMHA had IgG and IgM on erythrocytes, 5 had IgG, and 2 had IgM. Three horses had surface-bound IgG, including a horse with suspected penicillin-induced IMHA, a foal with neonatal isoerythrolysis, and a foal with clostridial septicemia. The DIF method was valuable in monitoring the response to therapy in the foal with neonatal isoerythrolysis.  相似文献   
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