中国枣果实病害研究进展

就20世纪80年代以后,我国枣果实生长发育及采后贮藏期病害研究进展进行了综述。采前枣果有9种病害,病原菌涉及到9个属真菌和4个属细菌。贮藏期枣果有4大类病害,但病原菌隶属12个属真菌。同一种病害不仅由单一病原菌侵染所致,而且往往是多种病原菌混合侵染引起的,因而给田间和贮期综合防治造成了一定困难。最后,提出了枣果病害研究上存在的主要问题和今后的研究方向。     

RT-PCR结合限制性内切酶分析法区分NDV强弱毒株

通过设计的特异引物，采用RT-PCR对1996年以来分离的45株国内新城疫病毒分离株进行鉴定和分析，并与其MDT实验结果进行比较，建立了RT-PCR结合BglⅠ内切酶分析区分新城疫强弱毒的方法。此方法操作简便、迅速，不仅能区分新城疫的强弱毒株，还可诊断AMPV-1引起的其他禽类的感染。     

靶向新城疫病毒NP基因的shRNA重组腺病毒表达载体的构建及鉴定

为验证腺病毒载体在鸡胚成纤维细胞(CEF)及鸡胚中递送小干扰RNA的可行性,本实验利用共转染技术将表达针对新城疫病毒(NDV)NP基因shRNA的重组质粒同源重组到pGSadeno腺病毒载体系统,用重组腺病毒感染CEF和鸡胚,通过红荧光报告基因的表达情况和荧光定量PCR检测NP基因mRNA的表达对其进行鉴定,并通过细胞形态观察、血凝价的测定评价其在CEF和鸡胚上对NDV增殖的影响。实验结果显示重组腺病毒能够感染CEF,感染CEF后6h、9h、12h与对照组比较NP基因mRNA的表达量分别降低了3.2倍,25.6倍,2.57倍,并能够推迟NDV致细胞病变效应,抑制NDV在鸡胚上的增殖,延缓鸡胚的死亡。本实验构建的重组腺病毒能够在CEF和鸡胚上递送特异的shRNA,为在CEF中的RNA干涉研究开辟了新的递送途径。     

不同佐剂新城疫疫苗对鸡体液免疫的影响

佐剂是疫苗研究的重要组成部分.选择合适的佐剂不仅可以增强疫苗的免疫效果,而且可以提高机体的抵抗力.不同佐剂对体液免疫会产生不同的作用.本研究选用白油佐剂、弗氏佐剂和蜂胶佐荆与新城疫病毒抗原混合制成疫苗,免疫雏鸡,探讨不同佐剂对鸡体液免疫的影响.结果表明,三种佐剂疫苗均有保护力,试验鸡饲养至61日龄均未发生新城疫,其中弗氏佐剂的效果最好,蜂胶佐剂的效果次之,油佐剂组检测到雏鸡61日龄时仍未出现抗体高峰值.     

新城疫病毒糖蛋白致病作用的研究

本实验以分子生物学和现代免疫学方法,对新城疫病毒（ＮＤＶ）中国毒株Ｆ４８Ｅ９和ＬａＳｏｔａ等毒株的糖蛋白的致病作用进行了探讨。结果表明,ＮＤＶ Ｆ和ＨＮ蛋白为糖蛋白,强弱毒株之间Ｆ蛋白裂解位点的氨基酸序列有明显的不同,并决定了毒株的毒力,但ＮＤＶ的致病作用尚需ＨＮ、Ｆ蛋白的协同作用,单一的Ｆ或ＨＮ蛋白不能构成ＮＤＶ的致病作用。     

Insect herbivores and pathogens of Alnus species in Uganda

Published documentation of insect pests and pathogens associated with Alnus species in Africa is very scarce. We surveyed damaging insects and pathogens, and arthropod natural enemies on Alnus acuminata and A. nepalensis in Kabale and Mbale districts, Uganda between March 1999 and August 2000 in order to identify the range and relative abundance of arthropods and pathogens associated with the Alnus species. Frequently encountered damaging insects on the Alnus species included Apis mellifera, Apion globulipenne, a Systates sp. (Coleoptera: Curculionidae), Phymateus viridipes, a Lobotrachelus sp. (Coleoptera: Curculionidae), Coloborrtics corticina and some Chrysomelidae. Some species such as Aphis fabae, Parastictococcus multispinosus and a Cacopsylla sp. (Homoptera: Psyllidae) were observed feeding on other agroforestry tree species and/or crops although they generally occurred at low population intensities. Spiders and parasitic Hymenoptera were the most common natural enemies. Diseases were more severe in nurseries than in the field. Damping-off caused by Fusarium oxysporum, Septoria brown leaf spot and stem canker were the most serious diseases of Alnus. The array of damaging insects and pathogens indicates a potential danger to the cultivation of Alnus species in Uganda as adoption of the species for agroforestry continues to expand in the country. In view of the increasing demand for Alnus species for agroforestry in Uganda, regular pest monitoring and appropriate control strategies are necessary.This revised version was published online in November 2005 with corrections to the Cover Date.     

Using participatory epidemiological techniques to estimate the relative incidence and impact on livelihoods of livestock diseases amongst nomadic pastoralists in Turkana South District, Kenya

A participatory epidemiological (PE) study was carried out with Turkana pastoralists in Turkana South District, Kenya, to determine the relative incidence of livestock diseasess and their impact on livelihoods. A sub-location was used as the sampling unit. A sub-location is the smallest administrative unit and is occupied by clusters of families (called adakars) that share common grazing patterns. A total of 32 sub-locations were randomly selected for the study. At least one focus group discussion involving more than 10 people was held with each adakar. In addition, key informant interviews involving local leaders and animal health service providers were conducted before or after the group sessions. PE techniques that were used with the stock owners include participatory mapping, relative incidence scoring, proportional piling, disease impact matrix scoring, seasonal calendars and probing. The methods used were pre-tested in four sub-locations that were excluded from further study. The study revealed that goats, with median score of 33 (10th and 90th percentiles of 25, 44, respectively) and sheep, median score of 20.5 (15, 26) were perceived to be the most abundant livestock species while goats (median score of 32 [21, 56]) and camels (median score of 22.5 [11, 33]) contributed the most to the livelihoods of the pastoralists. For goats, the overall relative incidence scores of peste des petits ruminants (PPR), contagious caprine pleuropneumonia (CCPP) and mange were 23.5% (15, 34), 25% (21, 45) and 20% (19, 28), respectively. The respective median scores for case fatality rates were 66% (45, 76.5), 62.5% (25, 100) and 73.2% (21.4, 85.7). Disease impact matrix scores indicated that mange was the most important disease of goats. Mange (range: 28–32%) and pox (range: 16–38%) were perceived to be the most prevalent diseases in camels. Livestock movements, limited access to veterinary services and stock theft were identified as key factors that contributed to the high prevalence and persistence of these diseases. This paper discusses strategies that could be used to control these diseases given the challenges associated with nomadic pastoralism and insecurity.     

A Droplet Digital PCR Method for Detection of Newcastle Disease Virus

This experiment was conducted to establish a droplet digital PCR (ddPCR) method for the detection of Newcastle disease virus (NDV). A ddPCR method was developed, which the primers and probes were designed based on the conservative regions of F gene of NDV. The concentration of primer and probe, the annealing temperature in ddPCR reaction were optimized. The sensitivity, specificity and reproducibility of ddPCR method were evaluated. In results, the optimal primer concentration and the probe concentration were 900 and 250 nmol·L^-1, the optimum annealing temperature was 55℃. The detection limit of ddPCR method was 1.8 copies·μL^-1 with a good linear response, it had no cross reaction with other six viruses (include IBV), the coefficient of variation of sample repetition was 2.4%. All the results showed that NDV ddPCR was sensitive and specific, it was suitable for quantitative detection of clinical samples infected with NDV.     

RT-PCR法快速鉴别新城疫强弱毒株的试验

通过对大量不同毒力NDV F基因核苷酸序列的分析，根据强弱毒株F0裂解位点的序列差异设计合成了三对引物，建立了快速诊断新城疫并能鉴别其强弱毒株的反转录-聚合酶链式反应(RT-PCR)方法，整个试验过程可在5h内完成。试验表明，该方法具有快速特异和操作简便的特点，不仅适用于对鸡胚毒的检测，而且适用于对病鸡组织匀浆液的检测，是新城疫鉴别诊断和流行病学调查的颇具潜力的分子诊断方法。