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81.
During early lactation, dairy cows may present a transient immunosuppressive state and develop anaplasmosis caused by Anaplasma marginale. In this study, clinical anaplasmosis in dairy cattle in the Thrace region of Turkey was investigated with respect to within-herd prevalence, vertical transmission, and genetic diversity. In March and September 2015, thirty lactating cows showed primary clinical signs of anaplasmosis, including fever, anaemia, decreased milk yield, anorexia, and laboured breathing. Symptoms disappeared in most cows after administration of long-acting oxytetracycline, but nine of them (30%) died. Following diagnosis based on clinical signs, microscopy and molecular findings, blood samples were collected from apparently healthy lactating cows (n = 184), pregnant heifers (n = 39) and newborn calves (n = 24). DNA was extracted from each sample and analyzed for the presence of major surface proteins (MSPs) of A. marginale, followed by sequencing to assess diversity of isolates. Microscopic examination of erythrocytes revealed A. marginale inclusion bodies in symptomatic cows. Examination of thin blood smears showed 3.8% of the lactating, clinically asymptomatic, cows to be infected with A. marginale, while nPCR detected 31.0% positive. A. marginale infection was not detected in pregnant heifers by either method. Congenital infection was found in one calf by nPCR. This is the first report of transplacental transmission of A. marginale in Turkey. The MSP4 sequence analyses showed high genetic diversity among the isolates, presenting 97.6-99.6% homology at the amino acid level. The sequences of MSP1a amplicons revealed genetic diversity providing three new tandem repeats. 相似文献
82.
Hepatozoonosis caused by Hepatozoon canis is an important tick-borne disease of dogs in tropical and sub-tropical regions throughout the world. In the present study evaluation of blood samples collected from 225 dogs presented at Small Animal Clinics, GADVASU, Ludhiana, Punjab (India) was done for the presence of H. canis by PCR based assay targeting a portion of 18S rRNA gene. Of the total samples subjected to PCR, an amplicon of 666 bp was detected in 13.78% samples whereas, routine blood smear examination revealed gamonts in 5.78% samples. Furthermore, prevalence of H. canis infection was found to be significantly associated with season, being highest in summer and lowest in winter while other risk factors e.g. age, sex and breed showed non-significant association. In terms of various clinico-pathological parameters, significant drop in haemoglobin, total red blood cell count, packed cell volume and lymphocytes were recorded in positive cases whereas the total white blood cell count was non-significantly increased. The haematological alterations in the positive cases were lymphopenia, anaemia, thrombocytopenia, relative neutrophilia, neutrophilic leucocytosis, eosinophilia, monocytosis and lymphocytosis while the biochemical profile revealed hypoproteinemia and increased levels of blood urea nitrogen and creatinine (in positive cases) pointing towards renal failure. 相似文献
83.
利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量 总被引:1,自引:0,他引:1
QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考. 相似文献
84.
This is the first study to investigate the prevalence and risk factors associated with Ehrlichia canis and Anaplasma platys positivity in dogs from Paraguay. Conventional PCR assays for the E. canis 16SrRNA gene and A. platys p44 gene were carried out in blood samples from 384 dogs from Asunción city, Paraguay. Sequencing and phylogenetic analysis were performed in selected positive E. canis and (16SrRNA gene) and A. platys (16S and p44 genes) samples. The overall prevalence of E. canis and A. platys in dogs in Paraguay was 10.41% (40/384) and 10.67% (41/384), respectively. Older dogs without veterinary care had higher odds for E. canis positivity and a higher number of dogs in the same household, as well as absence of anti-tick treatment were considered risk factors for A. platys. Ehrlichia canis and A. platys circulate in the dog population from Asunción, and are described for the first time in Paraguay. 相似文献
85.
不同的样本特性和提取方法对获得微生物总DNA的质量有重要影响。文章基于高含固率木质纤维素厌氧发酵物腐殖酸、酚类物质含量高、质地均一性差、微生物浓度低的特点,研究了4种方法提取不同高含固率粪秸厌氧发酵物中微生物总DNA的效果。结果表明,常规的十二烷基磺酸钠法(sodium dodecyl sulfate,SDS)、十二烷基磺酸钠和溴化十六烷基三甲铵结合法(sodium dodecyl sulfate and cetyltrimethyl ammonium bromide,SDS-CTAB)和商业的粪便试剂盒法提取的DNA质量均较差,SDS法和试剂盒法未能获得聚合酶链式反应(polymerase chain reaction,PCR)扩增目的条带,SDS-CTAB法得到的条带较模糊;改进SDS-CTAB法获得的DNA杂质少、纯度高,具有较好的稳定性,A260/A280和A260/A230值分别为1.74~1.86和1.65~1.86,每克样品的DNA浓度在50 ng·μL^-1以上,电泳条带单一齐整、清晰明亮,PCR扩增的目的条带清晰度高,适宜后续分子生物学技术的分析。林格氏液洗脱、聚乙烯吡咯烷酮-40(Polyvinyl Pyrrolidone-40,PVP-40)洗涤液除杂以及裂解液和多种酶联合破壁是改进SDS-CTAB法获得该类专一性样本高质量微生物总DNA的关键步骤。 相似文献
86.
以柑橘溃疡病菌特异性高的假定蛋白基因(登录号:AE008923.1)为靶标,设计并筛选4条引物来特异性识别靶标序列中的6个独立区域,建立柑橘溃疡病菌LAMP实时浊度检测方法。该方法利用实时浊度仪检测反应过程中产生的焦磷酸镁白色沉淀,实现对LAMP整个反应过程的实时监控。从镁离子浓度、甜菜碱浓度、dNTPs浓度、引物浓度和反应温度5个变量参数对 LAMP检测体系进行优化,并对方法的特异性、灵敏度、稳定性及实际样品检测效果进行评价。结果显示,该体系经优化后稳定性良好,可在66 ℃ 恒温条件下,60 min内完成检测;DNA检测下限可达0.02 ng/μL,灵敏度与荧光定量PCR方法相同,比常规PCR方法高1个数量级;能快速、准确地对田间疑似样品进行检测,与荧光定量PCR方法的检出情况一致,没有漏检。结果说明柑橘溃疡病菌实时浊度LAMP检测方法操作简便、所需时间短,特异性与灵敏度高,为柑橘溃疡病菌的快速筛查提供较好的途径。 相似文献
87.
以东北山樱桃(Cerasus sachalinensis Kom.)实生幼苗为试材,利用Hansatch PEA,采用叶绿素荧光诱导动力学理论和JIP-test数据分析方法,研究了对-羟基苯甲酸对叶片叶绿素荧光参数的影响。结果表明:幼苗在不同浓度对-羟基苯甲酸(0.1A、1A、10A)处理下的叶片光合性能指数(PIABS)、捕获的激子将电子传递到电子传递链中QA-下游的其他电子受体的概率(Ψo)、用于电子传递的量子产额(φEo)和单位面积有活性的反应中心数目(RC/CSm)较对照均有所下降;PSⅡ最大量子效率(φPo)、单位面积有活性的反应中心数目(RC/CS0)几乎没有变化;而PSⅡ受体侧的电子受体库容量(Sm)则高于对照,且呈"上升-下降"趋势。天线色素吸收的能量(ABS/RC)在低浓度下有小幅升高,反应中心捕获的能量(TRo/RC)、用于电子传递的能量(ETo/RC)及用于热耗散的能量(DIo/RC)则无明显变化。可见,对-羟基苯甲酸对东北山樱桃叶片光合结构的不同部位产生影响,进而降低光合性能,影响光合作用。 相似文献
88.
89.
S. Shanmugam K. H. Kjaer C.‐O. Ottosen E. Rosenqvist D. Kumari Sharma B. Wollenweber 《Journal of Agronomy and Crop Science》2013,199(5):340-350
This study analysed the alleviating effect of elevated CO2 on stress‐induced decreases in photosynthesis and changes in carbohydrate metabolism in two wheat cultivars (Triticum aestivum L.) of different origin. The plants were grown in ambient (400 μl l?1) and elevated (800 μl l?1) CO2 with a day/night temperature of 15/10 °C. At the growth stages of tillering, booting and anthesis, the plants were subjected to heat stress of 40 °C for three continuous days. Photosynthetic parameters, maximum quantum efficiency of photosystem II (PSII) photochemistry (Fv/Fm) and contents of pigments and carbohydrates in leaves were analysed before and during the stress treatments as well as after 1 day of recovery. Heat stress reduced PN and Fv/Fm in both wheat cultivars, but plants grown in elevated CO2 maintained higher PN and Fv/Fm in comparison with plants grown in ambient CO2. Heat stress reduced leaf chlorophyll contents and increased leaf sucrose contents in both cultivars grown at ambient and elevated CO2. The content of hexoses in the leaves increased mainly in the tolerant cultivar in response to the combination of elevated CO2 and heat stress. The results show that heat stress tolerance in wheat is related to cultivar origin, the phenological stage of the plants and can be alleviated by elevated CO2. This confirms the complex interrelation between environmental factors and genotypic traits that influence crop performance under various climatic stresses. 相似文献
90.
本文根据变量理论分布对区试中b值及CV值参数的数量分类,提出了适于目前普及推广的查表计算法。在此基础上,对区试品种提出了按主要生产性能指标以量定性的评定分类体系。从而推出可排除主观随意性干扰、较为完整严密的区试汇总模式,并以范例作了运用说明。 相似文献