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51.
To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg2+ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.  相似文献   
52.
为获得大熊猫犬瘟热病毒株,采集死亡大熊猫的心脏、肺、肝病料,研磨并反复冻融后收集上清液接种Vero细胞,待出现细胞病变(CPE)后,收集病毒液。用逆转录-聚合酶链反应(RT-PCR)鉴定病毒分离株,测定病毒TCID_(50),动物回归试验测定其毒力。结果显示,盲传到第5代时,Vero细胞出现圆缩、聚集、脱落等病变;PCR扩增出287bp片段,与预期相符;测序结果显示,该毒株与已发表的CDV SD(14)11毒株(亚洲-Ⅰ型)的同源性为98%;毒株的TCID_(50)为10~(-5.2)/mL;幼犬感染分离毒株后出现体温升高、鼻头发干、拉稀、眼鼻分泌出水样分泌物等症状。本研究成功分离出大熊猫源犬瘟热病毒株。  相似文献   
53.
对北海近岸养殖海域的3种弧菌(Vibrio)进行了分离鉴定,并对鉴定的弧菌进行耐药性分析。通过从北海近岸养殖海域随机采集水样,利用TCBS培养基对所采水样进行弧菌的分离和纯化,采用PCR方法和序列分析对弧菌进行鉴定,并对鉴定出的3种弧菌进行了常用抗菌药物耐药性的检测。共分离获得67株疑似弧菌菌株,经鉴定,19株为副溶血性弧菌(Vibrio parahaemolyticus),4株为霍乱弧菌(Vibrio cholera),4株为河流弧菌(Vibrio fluvialis)。耐药性分析研究表明,19株副溶血性弧菌总体显示对10种常用抗菌药物具有耐药性,其中100%的菌株对四环素、卡那霉素和红霉素表现耐药,94.7%的菌株对氨苄西林耐药、84.1%的菌株对左氧氟沙星和78.9%的菌株对环丙沙星耐药;4株霍乱弧菌和4株河流弧菌对10种抗菌药物的耐药性均较强,但敏感度低,两者对复方新诺明均表现为中介敏感。本研究提示,北海近岸养殖海域中弧菌种类较多,其中以副溶血性弧菌为主,大多对常用抗菌药物具有耐药性,对北海近岸海水养殖疾病防治具有一定的指导意义。  相似文献   
54.
Comparison of sampling sites and detection methods for Haemophilus parasuis   总被引:3,自引:0,他引:3  
Objective To improve the isolation rate and identification procedures for Haemophilus parasuis from pig tissues. Design Thirteen sampling sites and up to three methods were used to confirm the presence of H. parasuis in pigs after experimental challenge. Procedure Colostrum‐deprived, naturally farrowed pigs were challenged intratracheally with H parasuis serovar 12 or 4. Samples taken during necropsy were either inoculated onto culture plates, processed directly for PCR or enriched prior to being processed for PCR. The recovery of H parasuis from different sampling sites and using different sampling methods was compared for each serovar. Results H parasuis was recovered from several sample sites for all serovar 12 challenged pigs, while the trachea was the only positive site for all pigs following serovar 4 challenge. The method of solid medium culture of swabs, and confirmation of the identity of cultured bacteria by PCR, resulted in 38% and 14% more positive results on a site basis for serovars 12 and 4, retrospectively, than direct PCR on the swabs. This difference was significant in the serovar 12 challenge. Conclusion Conventional culture proved to be more effective in detecting H parasuis than direct PCR or PCR on enrichment broths. For subacute (serovar 4) infections, the most successful sites for culture or direct PCR were pleural fluid, peritoneal fibrin and fluid, lung and pericardial fluid. For acute (serovar 12) infections, the best sites were lung, heart blood, affected joints and brain. The methodologies and key sampling sites identified in this study will enable improved isolation of H parasuis and aid the diagnosis of Glässer's disease.  相似文献   
55.
Brown stripe disease is a severe foliar fungal disease of sugarcane worldwide and is widespread in all sugarcane planting areas in China. Brown stripe is a major disease that seriously affects the output and quality of the sugarcane industry in Yunnan Province, China's second-largest sugar base, while the pathogen of this disease remains not yet fully understood. To address this, we isolated and identified the fungi associated with 68 leaf samples showing typical symptoms of brown stripe from 22 sugarcane varieties in different areas of Yunnan Province. A total of 113 isolates were obtained, which were morphologically similar. Of these, 64 representative isolates were sequenced for the internal transcribed spacer region (ITS), GAPDH and EF-1α loci. All representative isolates grouped with the type strain of Bipolaris setariae in the phylogenetic trees inferred with individual and concatenated sequences of ITS, GAPDH and EF1-α. Pathogenicity test results showed that B. setariae strains were able to induce typical symptoms of brown stripe. The results obtained in this study clarify that only B. setariae is associated with sugarcane brown stripe in Yunnan, China. It is recorded here for the first time as a pathogen causing sugarcane brown stripe in Yunnan, and it is able to infect many major cultivars and new varieties, posing a new threat to the sugar industry in Yunnan Province. In addition, these results provide the scientific basis for the future breeding of disease-resistant varieties and effective prevention and control of sugarcane brown stripe disease.  相似文献   
56.
The dynamic characteristic of active isolation atuator on flexible foundations was studied under the installation of parallel structure and foundation cancellation.The influencing relationships between active isolation force of the different installation ways and passive isolation system and flexible foundation were analyzed and compared.Scientific basis for selecting installation ways of active isolator is provided in this paper.  相似文献   
57.
不同强度温度刺激对棉花黄萎病的影响   总被引:5,自引:0,他引:5  
采用三个不同抗性的棉花品种、三个不同的黄萎病菌系和三个温度梯度,完全随机区组设计,在人工气候箱中研究刺激不同时间对棉花黄萎病发病的影响。结果表明,刺激温度的高低和品种的抗性对棉花黄萎病的发生与扩展有显著的影响,而菌系致病力以及刺激时间长短对棉花黄萎病的发生也存在显著差异,但影响较小;品种的抗病性对棉花黄萎病的发生起着至关重要的制约作用;温度对黄萎病的发生与扩展起着关键作用。初步明确了影响棉花黄萎病发生与扩展的几个因子间的互作效应及其影响程度。  相似文献   
58.
灵芝孢子食用方法与破壁技术研究进展   总被引:8,自引:0,他引:8  
为了促进灵芝孢子破壁技术的应用 ,对目前普遍运用的 5类破壁技术和方法进行了对比分析 ,表明破壁后的灵芝孢子更易被人体消化吸收 ,说明灵芝孢子的破壁后提取生理有效成分已经成为今后发展的方向  相似文献   
59.
对橡胶悬架装置动特性的改进探讨   总被引:1,自引:0,他引:1  
以橡胶悬架装置为研究对象,考虑其动态特性与高速车辆动力总成低频与高频要求不同之间的矛盾,试图通过改进橡胶悬架装置的结构提高其动特性。建立了改进结构的橡胶悬架装置的力学模型,并对其进行了理论分析和仿真计算,结果表明改进后的橡胶悬架装置的动特性有了显著提高,对我国橡胶悬架装置的结构改进具有借鉴和指导意义。  相似文献   
60.
以大理州"红阳"猕猴桃枝干为试材,采用田间调查法调查病原菌对其危害状况,对采集的10份病样采用常规组织分离法分离病原,单孢纯化法获得病原菌,采用形态和分子法鉴定病原种类,以期明确大理州"红阳"猕猴桃枝干病害的发生状况及危害病原。结果表明:大理州"红阳"猕猴桃枝干病害发生严重,病株率高达69.6%,5个采样点共分离得到15个菌株,其中以链格孢菌(Alternaria alternata)、厚坦镰刀菌(Fusarium chlamydosporum)、小新壳梭孢菌(Neofusicoccum parvum)和间座壳属中一种(Diaporthe bohemiae)共4类病原菌出现频率最高,是优势种,分离率分别为55.6%、52.4%、49.8%和50.1%。利用柯赫氏法则对分离所得的所有菌株进行致病性测定,证实了优势菌株对猕猴桃茎杆的致病性,进一步明确了引起大理州猕猴桃茎杆的病原真菌。  相似文献   
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