茶多酚对高酒糟日粮肉鸡生产性能和抗氧化特性的影响

本研究旨在评估添加茶多酚对高酒糟(distillers dried grain with solubles DDGS)日粮肉鸡生产性能和抗氧化特性的影响。将400只21日龄肉鸡随机分为5个处理,每个处理4个重复,每个重复20只。正对照组为玉米-豆粕型日粮,负对照组为玉米-豆粕-DDGS型日粮(15%DDGS),处理组分别在负对照日粮中添加100、300、500 mg/kg茶多酚。结果表明,与正对照组相比,后期日粮中添加15%DDGS对肉鸡生产性能无显著影响(P>0.05),但血液的GSH-Px降低8.31%(P<0.01),胸腿肉T-AOC含量降低,腿肉MDA值显著升高(P<0.05)。添加茶多酚提高了血液的T-AOC和GSH-Px活性(P<0.05)。随着茶多酚添加剂量的升高,T-AOC和GSH-Px有升高的趋势,但各水平组之间差异不显著;茶多酚添加300 mg/kg以上能显著降低胸腿肌MDA值(P<0.05),提高T-AOC含量,改善肉质的抗氧化特性。本研究结果表明,在肉鸡高DDGS日粮中添加茶多酚可以改善肉鸡机体及肉质的抗氧化特性。     

单细胞转录组测序技术及其在肌内脂肪细胞研究中的应用

单细胞转录组测序(single-cell RNA sequencing, scRNA-seq)技术在单细胞水平对转录组进行测序,可从表达量、细胞量及细胞组成等多种角度描述样本,主要包括单细胞分离、mRNA反转录、文库构建、转录组测序和数据分析等步骤。单细胞分离是scRNA-seq技术操作的第一步,常用的细胞分离方法有连续稀释法、显微操作法、荧光激活流式细胞术、激光捕获显微切割术和微流控技术。基于不同细胞捕获、cDNA扩增和文库构建开发了许多scRNA-seq方法,如CEL-seq2、Drop-seq、MARS-seq、SCRB-seq、Smart-seq、Smart-seq2等。与传统测序方法相比,scRNA-seq可识别单个细胞的基因表达信息、记录细胞的空间位置、跟踪不同细胞谱系在分化过程中的轨迹,这为研究难以大量提取的肌内脂肪细胞分子特异性和发生分化过程带来极大便利。作者简要介绍了scRNA-seq技术的关键步骤,比较分析了单细胞分离和转录组测序方法的优缺点,综述了肌内脂肪细胞的起源以及scRNA-seq在肌内脂肪细胞的亚群和标记基因鉴定以及细胞分化轨迹追踪中的应用,以期为肌内脂肪...     

两种洗涤方法对马传染琼扩抗原生产试验

用两种不同洗涤方法生产马传染琼扩抗原。试验结果表明,用硫酸重铬酸钾清洗液洗刷的组织培养克氏瓶,细胞生长旺介线清楚,立体感强,接种马传染性贫血病毒后。细胞抗原产量高于用清洁剂刷的克氏瓶培养的细胞抗原的２￣３倍。     

Distribution and characterization of class 1 integrons in Salmonella enterica serotype Gallinarum biotype Gallinarum

Fowl typhoid caused by Salmonella enterica subsp. enterica serotype Gallinarum biotype Gallinarum is the most important chicken disease in Korea. Due to appearance of new or multiple antibiotics resistances in the recently isolated strains, it was difficult to control the disease using antibiotics in our country. Therefore, the prevalence and genetic contents of class 1 integrons in biotype Gallinarum isolated between 1992 and 2001 were investigated by PCR and direct sequencing, respectively. Out of 90 strains, 35 (39%) carried class 1 integrons. The 1.0, 1.6 and 2.0kbp amplicons were amplified in 32 strains (36%), 2 strains (2%) and 1 strain (1%), respectively. The 1.0, 1.6 and 2.0 kbp amplicons contained one (aadA1a), two (aadB-aadA1b) and three cassettes (dhfrXII-orfF-aadA2), respectively, providing resistances against aminoglycosides (aadA1a, aadA1b, aadB, and aadA2) and trimethoprim (dhfrXII). The integron-carrying strains of biotype Gallinarum appeared in 1996 and acquired additional cassettes in 2000. Although the resistances to ampicillin, tetracycline and chloramphenicol are unrelated to class 1 integrons, relatively high prevalence of integron in biotype Gallinarum may be a dormant threat to the chemotherapy of the disease in the near future because of potency to acquire additional antibiotics resistances.     

小反刍兽疫病毒M基因截短表达及鉴定

目的将小反刍兽疫病毒M蛋白基因截短表达后用于特异性单抗制备及临床抗体水平检测。方法:用在线分析软件BepiPred分析小反刍兽疫病毒M蛋白潜在的B细胞线性表位,以本实验室构建的pCR2.1T-PPRV M质粒为模板,扩增三段截短的M基因,纯化后的PCR产物分别与克隆载体pCR2.1T连接,筛选出的阳性重组质粒经双酶切后,分别与表达载体pET-32a(+)及pGEX-6p-1连接,再将鉴定为阳性的重组质粒转化入E.coli BL21(DE3)菌株诱导表达,并用SDS-PAGE及Western blot验证。结果 PCR产物电泳,得到与预期大小相符的特异性片段。对连接克隆载体及表达载体的重组质粒双酶切后,均出现与预期一致的片段,DNA测序表明,插入片段序列与小反刍兽疫Nigeria75/1株M蛋白基因完全一致。重组蛋白经SDS-PAGE及Western blot鉴定,证明所构建的重组蛋白均获得高效表达,并均具有良好的反应原性。结论成功表达了小反刍兽疫截短M基因的蛋白,为制备特异性单抗及小反刍兽疫抗体检测奠定了一定基础。     

Small molecule lactoferrin with an inflammatory effect but no apparent antibacterial activity in mastitic mammary gland secretion

We have identified various lactoferrin (Lf) molecules in mastitic mammary gland secretions (MGSs), and these Lf molecules were examined for their physiological function in MG. These Lf molecules were isolated by Con A affinity chromatography, and then analyzed by various electrophoresis methods and N-terminal amino acid sequencing. The low Con A affinity Lf was found to have low molecular peptides as compared with the 86 kDa of the high Con A affinity Lf, which is usually detected in healthy MGSs. The N-terminal amino acid sequence of each of the small molecular Lfs were confirmed as fragments of 86 kDa Lf. This low Con A affinity Lf stimulated spleen adherent cells to produce more O(2)(-) than 86 kDa Lf. Furthermore, the low Con A affinity Lf showed low antibacterial activity against E. coli and S. aureus, and had decreased iron-binding capacity in comparison with 86 kDa Lf. Moreover, the 86 kDa Lf could stimulate bovine T cells or macrophages to produce IFN-gamma, IL-4, IL-6, and IL-1alpha. However low Con A affinity Lf induced the production of TNFalpha, but not physiological T cell or macrophage cytokines. It was also found that when the healthy MGs of dry cows were injected with the low Con A affinity Lf, there was an increase in polymorphonuclear cells together with TNFalpha, MCP-1, and IL-8 production. These results suggested that low Con A affinity Lf in mastitic MGSs differed from 86 kDa Lf in physiological characteristics, and, that it induced an inflammatory reaction in MGs.     

影响冻干乳酸菌存活性的相关因素

本文综述冻干乳酸菌发酵剂制备中相关因素对冷冻干燥和贮藏期间乳酸菌存活率的影响，包括内在因素、生长因素、非致死性处理、干燥介质、贮藏和复水等方面因素。     

复方罗红霉素可溶性粉对鸡葡萄球菌病的疗效试验

采用人工诱发的病理模型,按推荐剂量治疗,验证复方罗红霉素可溶性粉对鸡(试验时30日龄)葡萄球菌病的疗效.试验结果表明,复方罗红霉素可溶性粉对鸡葡萄球菌病有很好的治疗效果,有效率达96.7%;推荐剂量治疗组的死亡率极显著地低于感染对照组(P<0.01);推荐剂量治疗组与健康对照组的增重差异不显著(P>0.05),但极显著高于感染对照组(P<0.01).     

豆科牧草追施钾肥试验

试验结果表明，当旱地红壤养分中的速效钾含量在７５．５ｐｐｍ情况下，白三叶每公顷追施氯化钾２２５ｋｇ，结果施肥比对照每公顷增产鲜草７７７９ｋｇ，每公顷氯化立鲜草３４．６ｋｇ，增产率达２９％。按生物统计ｔ测验，增产显著。当稻田速效钾含量在４５ｐｐｍ条件下，给紫云英每公顷施用氯化钾１１２．５ｋｇ，结果施肥比对照每公顷增产鲜草５０００－７７５０ｋｇ，平均每公顷增产６７５０ｋｇ。每公斤钾肥可增产粮食２３．５     

Immune responses of orange-spotted grouper,Epinephelus coioides,against virus-like particles of betanodavirus produced in Escherichia coli

Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15 μg g^?1 fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 μg g^?1 FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12 h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper.