马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。     

以CP23重组蛋白为抗原建立检测微小隐孢子虫抗体间接ELISA方法的研究

摘要：以在E.coli高效表达的微小隐孢子虫子孢子表面抗原CP23为抗原，以辣根过氧化物酶(HRP)标记的山羊抗鼠IgG为二抗，建立了检测微小隐孢子虫抗体的间接ELISA方法。经检测筛选出最佳反应条件为1μg／孔纯化的E.coli表达的CP23抗原包被酶标板，用10％免血清进行封闭，以正常E.coli裂解上清液稀释待检血清。实验表明应用CP23重组蛋白作为诊断C.parvum抗原具有特异性高、抗原易纯化和成本低等特点。     

Food intake and energy expenditure in growing cats with and without a predisposition to overweight

Overweight and obesity are multifactorial diseases caused by an imbalance in energy metabolism. An underlying genetic predisposition is often a factor in these conditions. In the cat breeding family of the Institute of Animal Nutrition at the Vetsuisse Faculty, University of Zurich, a segregating overweight phenotype with a genetic contribution was observed. From this breeding family, 26 kittens were followed from birth up to 8 months of age. During this time, food intake was measured using an automatic feeding station, and energy expenditure was investigated using indirect calorimetry at the ages of 4 and 6 months. Dual‐energy X‐ray absorptiometry (DEXA) was performed and blood glucose, leptin and insulin were measured at the ages of 4, 6 and 8 months. The kittens were also weighed daily for the first 2 weeks of life, every second day until weaning and once per week until 8 months of age. The body condition score (BCS) was evaluated monthly between 2 and 8 months of age. The main finding of this study is that a predisposition to overweight is connected to a higher food intake early in life, with no significant alterations in energy expenditure. The leptin blood levels were related to body fat percentage, and insulin sensitivity did not seem to be affected.     

检测马疱疹病毒4型抗体间接ELISA方法的建立及应用简

In order to establish an indirect ELISA method for detection of equine herpesviruses type 4 （EHV4） antibody, the glycoprotein G （gG） protein with specific epitope worked as a detection antigen. After optimizing conditions, the indirect ELISA method was developed successfully，specificity and repeatability tests were determined. The result was that the EHV4 gG only reacted with antibodies against EHV4;but not with antibodies against EHV1; both the intro-batch and inter-batch variation coefficiencies were lower than 10%.Concordance of the indirect ELISA relative to commercial EHV1/4 antibody kit was above 90%.The results indicated that the indirect ELISA method could be used for the detection and epidemiological surveys of EHV4 infection.     

产蛋期蛋鸡不同类型玉米净能研究

本试验利用禽用开放式呼吸测热装置进行能量代谢试验,通过间接测热法结合替代法测定不同类型玉米在产蛋期蛋鸡饲粮中的表观代谢能和净能。选用34周龄产蛋期海兰褐蛋鸡180只,随机分为6组,每组30只。试验选用1种玉米-豆粕型基础饲粮和5种待测饲粮。待测饲粮由5种待测玉米(3种2018年10月收获的正常玉米和2种2016年收获储存3年的陈化玉米),分别以50%比例替代基础饲粮构成。试验鸡在舍内笼养,预试期7 d,正试期27 d,其中正试期分为3期,每期9 d(适应3 d、呼吸测热3 d、绝食测热3 d)。每期试验中,从每组中选择4只试验鸡,称重后分别放入呼吸测热装置的12个代谢室(每个代谢室2只),每2个代谢室对应1种饲粮,测定气体交换和排泄物总量,呼吸测热的同时进行消化代谢试验。结果表明:与基础饲粮相比,5种玉米待测饲粮的表观代谢能显著提高(P<0.05),3种正常玉米待测饲粮的净能显著高于基础饲粮和2种陈化玉米待测饲粮(P<0.05);2种陈化玉米的表观代谢能和净能显著低于3种正常玉米(P<0.05)。本试验中,3种正常玉米的表观代谢能分别为16.19、15.85、16.17 MJ/kg,2种陈化玉米的表观代谢能分别为15.12和15.06 MJ/kg;3种正常玉米的净能分别为12.39、12.57、12.25 MJ/kg,2种陈化玉米的净能分别为11.29和12.05 MJ/kg。     

间接免疫荧光染色检测鸭源副黏病毒及在鸭体内的抗原定位

以抗新城疫病毒F蛋白的单克隆抗体为一抗,建立了间接免疫荧光染色法(IFA)检测石蜡切片中鸭副黏病毒(DPMV)的方法。以建立的IFA对DPMV人工感染鸭的各组织器官进行检测,结果显示:各组织器官均能检测到DPMV,但不同时间取样,阳性信号分布的器官不同。脾脏、胸腺、法氏囊、肠道、肝脏、肺脏DPMV的阳性检出率较高,表明这些器官为DPMV的主要靶器官。在所检测的阳性组织中,病毒抗原分布在细胞浆中。IFA检测石蜡切片中的DPMV具有直观、特异性强的优点,是对DPMV进行检测和抗原定位的良好方法。     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

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.     

Sentinel lymph node mapping in canine mast cell tumours using a preoperative radiographic indirect lymphography: Technique description and results in 138 cases

Several sentinel lymph node (SLN) mapping techniques, to detect nodal metastasis in canine tumours have been investigated in the last 10 years in veterinary oncology. The purpose of this prospective study was to describe a reliable, quick, and inexpensive technique for SLN mapping in canine patients affected by cutaneous and subcutaneous mast cell tumours (MCT). Eighty dogs were enrolled in this study for a total of 138 cytologically diagnosed MCTs. Sentinel lymph node mapping was performed by injecting iomeprole peritumorally followed by serial radiographs at 1, 3, 6 and 9-min post injection. A total of 168 SLNs were detected, 90% at first radiograph, 1 min after the peritumoral iomeprole injection, while in the rest of the cases SLN was identified at 3 min. Sentinel lymph nodes detected by the preoperative radiographic indirect lymphography with iomeprole (PRILI) differed from regional lymph nodes in 57% of cases. The PRILI technique detected simultaneously multiple SLNs in the 26% of cases and multiple lymph centers in the 31% of MCTs. To allow the surgical identification of the SLNs, a peritumoral injection of methylene blue was performed at the time of surgery. This study reports a widely available technique for SLN mapping using digital radiographs in combination with a water-soluble medium, representing a cost-effective alternative to other SLN mapping procedures. Based on our results, this technique can be effective for SLNs mapping in dogs with MCTs but further comparative studies are needed to assess its reliability and efficacy in different tumours.     

间接地下滴灌导水装置规格参数模型研究

根据灌水过程中质量守恒原理建立了导水装置规格(直径,高度)参数求解模型,该模型中综合考虑了滴头流量、灌水定额、土壤水力特性参数以及活动水位等因素对导水装置规格的影响。通过室内恒定水头积水入渗试验的设计和研究,定量化了土壤水力特性参数对导水装置规格参数模型的作用,并研究了导水装置内水位深度的动态变化规律。结果表明,当忽略导水装置内积水深度的动态变化对土壤水力特性参数的影响时,与一维垂直入渗规律相似,导水装置内积水入渗累计入渗量与时间的关系可用菲利普公式进行拟合,但在土壤类型一定时,间接地下滴灌的参数渗吸系数S和稳定入渗速率if还与土壤上方的积水面积有关。当滴头流量大于土壤入渗速率时,导水装置内开始产生积水,且积水深度处于不断上升的过程。该模型的建立为间接地下滴灌导水装置规格的选取提供了理论指导。