东方田鼠肝脏T7噬菌体展示cDNA文库的构建

本项研究的目的是构建东方田鼠肝脏T7噬菌体展示cDNA文库,为筛选东方田鼠抗血吸虫病抗性相关基因奠定基础.用TRIzol试剂提取东方田鼠肝脏总RNA,分离纯化mRNA,经反转录合成双链cDNA.在双链cDNA末端加上EcoRⅠ/HindⅢ定向接头并用EcoRⅠ和Hind Ⅲ酶切,使其两端分别带EcoRⅠ和HindⅢ粘性末端.用Mini Column纯化、收集300 bp以上的双链cDNA片段,再连接于带有EcoRⅠ和HindⅢ末端的T7 Select 10-3b载体,经体外包装后,以BLT5403为受体菌构建T7噬菌体展示cDNA文库.经测定,库容量为1.3×107 PFU/mL,扩增后文库滴度为1.8×1011 PFU/mL.对从原始文库中随机挑取的100个噬菌斑进行PCR鉴定,重组率为91.7%,阳性克隆片段大小分布在200 bp～1 000 bp,其中有95.5%的插入片段大于300 bp.用日本血吸虫童虫可溶性抗原对文库进行了初筛,得到了21个ESTs,将这些阳性噬菌体克隆和血吸虫童虫共培养,其中大部分克隆诱导的童虫死亡率比阴性噬菌体对照高出2%～13%.     

互花米草生境中人工巢巢捕食研究

为了研究江苏大丰麋鹿国家级自然保护区互花米草生境中繁殖鸟巢的被捕食情况及影响捕食的因素,2014年3~6月,在保护区第三核心区鸟类繁殖生境中,使用人工巢及真实鹌鹑蛋进行巢捕食实验的方法进行巢捕食实验,分析不同巢高度、覆盖度、边缘距离、植被密度等因素对鸟巢被捕食的影响。实验共放置520个人工巢,1 040枚卵,使用红外相机对部分人工巢进行监测。结果表明:在江苏大丰麋鹿自然保护区互花米草生境内,人工巢存在较高的被捕食率,共有352巢被捕食,占67.7%,对11个变量进行逻辑斯蒂回归,巢高度、植被均高、主要植被密度、距边缘距离、水平覆盖度、上层垂直覆盖度等6个变量进人最终模型,巢高度、距边缘距离与巢被捕食显著相关。通过红外相机拍摄从102个监测巢中记录到69个捕食事件,68巢黑线姬鼠(Apodemus agrarius)和1巢黑眉锦蛇(Elaphe taeniura)捕食。我们认为互花米草生境中缺乏啮齿类动物的天敌是当地人工巢被捕食的主要原因。     

北京地区早熟禾属植物种质资源研究

为收集优良的草坪种质资源,对北京地区野生早熟禾属(Poa L.)植物资源进行了实地考察。北京共有早熟禾属植物20种1个亚种,其中7个种及1个亚种为新分布,西部和北部山区早熟禾属植物物种分布较为丰富,尤其是东灵山、百花山、海坨山及喇叭沟门。从垂直分布看,草地早熟禾(Poa pratensis L.)为广布种,从平原到高海拔地区均有分布。硬质早熟禾(Poasphondylodes Trin.)在低山区常见,其他物种主要分布在海拔1000 m以上的地区。通过研究23份来自不同居群的草地早熟禾花序特征,发现不同居群的花序性状间存在较大的变异,花序基部最短分枝长的变异最大,花序小穗数次之。     

应用发光细菌法检测饲用抗生素单一及联合毒性的研究

以HgCl2为毒性参照,以等毒性配比法研究了饲用盐酸金霉素、吉他霉素、盐霉素和黄霉素对发光细菌的单一及联合毒性。结果表明:盐酸金霉素、吉他霉素、盐霉素、黄霉素对明亮发光杆菌的急性毒性EC50分别为9.21、146.13、4.24 mg/L和134.68 mg/L,发光细菌的相对发光强度(RLU)随着抗生素浓度的提高而降低,并分别在2~16、25~200、1~8 mg/L和25~200 mg/L范围内呈良好线性关系;多元饲用抗生素对发光细菌具有联合毒性的作用,吉他霉素与盐酸金霉素、盐霉素和黄霉素3种抗生素的二元组合对发光细菌的联合毒性表现为较强的协同作用,而盐酸金霉素、盐霉素和黄霉素两两分别组合对发光细菌的联合毒性表现为拮抗作用;三元及四元抗生素混合体系对发光细菌的联合毒性均表现为拮抗作用。     

寄生虫半胱氨酸蛋白酶核酸疫苗的研究进展与应用前景

就寄生虫半胱氨酸蛋白酶的一些生理生化与免疫学性质进行了概括,阐述了该酶与宿主免疫应答之间的作用关系,证明了该酶具有作为研制高效核酸疫苗的条件,同时综述了寄生虫半胱氨酸蛋白酶核酸疫苗的发展以及应用前景。     

Effects of Feeding Different Dietary Crude Protein and Digestible Energy to Yili Mares during Late Lactation Stage on Growth Development and Blood Biochemical Parameters of Foals

In this paper, thirty Yili mares aged 11 to 12 years with weighing 410 kg ± 30 kg, parity of 4 to 5 in third lactation month were selected, and were randomly divided into six groups, each group with five repeats.Two factors quadratic regression orthogonal rotational combination design was adopted to evaluate the effects of feeding different dietary CP and DE to mares on growth development and blood biochemical parameters of foals.Daily DE and CP feeding levels of the 6 groups were 92 MJ/d and 1.20 kg/d, 125 MJ/d and 1.20 kg/d, 92 MJ/d and 1.55 kg/d, 107 MJ/d and 1.35 kg/d, 125 MJ/d and 1.44 kg/d, 115 MJ/d and 1.55 kg/d, respectively.The results showed that body weight, body height, chest circumference and cannon circumference of foals of four and five months were not significant affected by maternal protein and energy intake (P>0.05).As compared with ration contained high DE (125 MJ/d) and low CP (1.20 kg/d), feeding with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) improved body length of foals of four and five months significantly (P<0.05).From the blood biochemical parameters, such as total protein, albumin, triglyceride, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, urea nitrogen, creatinine, alanine aminotransferase, there were no significant differences among all groups (P>0.05).Mares fed with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) obviously improved the aspartate aminotransferase and uric acid level.In summary, Yili mares during lactation late stage fed with 1.35 kg/d CP and 107 MJ/d DE daily could significantly improve body length development of foals, and foals nitrogen metabolism was altered by maternal dietary nutrition through increasing blood aspartate aminotransferase activity.     

Effects of Donor Cells Treated with Egg Extracts on the Development Competence of Cloned Embryos

Japanese Balck cattle fetal fibroblasts (JBCFF) were induced with Xenopus leavis egg extracts and somatic cell nuclear transfer (SCNT) was carried out with the reprogrammed JBCFF as donor cells in order to investigate their effects on SCNT efficiency.Three samples of egg extracts were acquired from different Xenopus laevis.The protein contents and kinds in extracts were evaluated with BCA Protein Quantification Kit and SDS-PAGE.Concentration of Digitonin to permeabilize JBCFF was optimized and assessed with PI staining.Reprogrammed cells treated with egg extract were used as donor in SCNT.Additionally the reconstructed embryos were activated with ionomycin+6-DMAP and A23187+6-DMAP to compare their effects on the development competence.The protein contents of extracts samples were 56.2255,64.6570 and 71.2158 μg/mL,respectively,the each extract had the same composition about 40-55 and 70-100 ku.The optimal concentration of Digitonin was 7 μg/mL and the permeabilization rate was 55.44%.After extracts treatment and continuous culture for 6-7 d,JBCFF formed well-defined colony structures.No significant composition difference was found in rates of fusion (92.83% vs 96.04%),cleavage (89.64% vs 89.78%) and blastocyst formation (24.06% vs 23.12%) of cloned embryos when the colony cells and JBCFF without extracts treatment were used as donor cells (P>0.05).Similarly,the two activation methods had no significant effect on the developmental competence of cloned embryos (cleavage rate 92.16% vs 92.28%,blastocyst rate 23.21% vs 24.18%).Conclusively,Xenopus leavis,egg extracts could induce JBCFF reprogramming to a low differentiated state.However donor cells with reprogramming partially could not improve the development of cloned embryos and its mechanism requires further research.     

Effect of AIR on Inflammatory Reaction Infected by Brucella

In order to investigate the effect of AIR on inflammatory reaction infected by Brucellamelitensis (16M), the AIR domain of Tecpr1 gene of murine macrophages RAW264.7 were knocked down (I-A), overexpressed (O-A) and reversed (OA-IA). Using the chlorine fluorescein (DCFH-DA) as a probe, we detected the variation of ROS production and mitochondria distribution by confocal laser scanning microscopy. We observed the expression changes of NLRP3, ASC and Caspase-1 by qRT-PCR and the expression changes of IL-18,IL-1β and Caspase-1 in host cells by ELISA. The results showed that 16M could stimulate RAW264.7 cells to produce ROS by time-dependent pathway, and I-A group and O-A group showed more abnormal accumulation of mitochondrial. The results of qRT-PCR and ELISA suggested that it had effect on the expression levels of NLRP3, ASC,Caspase-1 and IL-18, IL-1β and Caspase-1 in cells of different groups. Those results indicated that with AIR gene deletion, the release amount of ROS changed, mitochondrial clustered abnormally, and AIR was closely related to the activation of inflammasomes and induction of inflammatory reactions.     

花椒叶浸提液对紫云英种子的化感作用

研究了不同浓度(5,10,20,40 g/L)的花椒叶浸提液对供试植物紫云英的种子萌发、幼苗根长及生理生化的影响,探究花椒化感作用的可能机理,从而为全面了解花椒林草间作种间关系及为农林复合经营和科学栽培提供理论依据.结果表明:1)花椒叶浸提液对紫云英种子萌发和幼苗根长均具有不同程度的抑制作用,且作用强度与浓度呈正相关(...     

不同旋毛虫隔离种在猪体内的分布

本文研究了黑龙江省猪旋毛虫和国际标准株旋毛形线虫（Trichinella spiralis)在猪体内的分布情况，结果表明，猪旋毛虫和T.spiralis在猪体内寄生寄生密度较大的部位是：膈肌、舌肌、咬肌、腓肠肌、肩胛肌、肋间肌、颈肌、前腿肌，臀肌和背最长肌寄生密度较小。