稳定表达猪圆环病毒2型Cap蛋白的CHO-K1细胞系的建立及免疫原性分析

建立高表达PCV2-Cap蛋白的CHO-K1悬浮稳转细胞系,鉴定蛋白的免疫原性,为开发新型且有效防治猪圆环病毒的亚单位疫苗奠定基础。构建重组质粒pEE12.4-PCV2-Cap,转染CHO-K1细胞,通过加压筛选,有限稀释,细胞悬浮驯化及Western blot检测得到高表达Cap蛋白的悬浮稳转单克隆细胞株,并对该细胞株进行发酵,纯化得到的目的蛋白,通过小鼠免疫验证其免疫原性。结果表明PCV2-Cap蛋白能够在CHO-K1细胞中正确表达;发酵过程中活细胞密度高达6×10~6个·mL^-1,细胞活力在80%以上,PCV2-Cap蛋白表达量约为370 mg·L^-1;利用间接ELISA检测小鼠免疫后血清中的抗体水平,证明了利用CHO-K1细胞生产的Cap蛋白具备良好的免疫原性。本研究成功构建了表达PCV2-Cap蛋白的CHO-K1悬浮稳转细胞系,并进一步对目的蛋白进行免疫原性分析,为猪圆环病毒亚单位疫苗的开发打下扎实的基础。     

银狐垂体细胞系构建研究

分别采用组织块法和消化法2种接种方法进行银狐垂体细胞系构建研究。在消化法中使用不同浓度的胰蛋白酶来检验效果,同时在原代培养期使用反复差速贴壁法纯化,传代培养期结合使用反复差速贴壁以及两步消化法进行处理。结果表明：组织块法接种后成纤维细胞生长明显,原代培养后期成纤维细胞已为主要细胞;降低胰蛋白酶浓度能降低对细胞损伤,但消化时间会增长;采用消化法接种在抑制成纤维细胞生长方面效果好于组织块法。纯化效果上,结合反复差速贴壁法与两步消化法处理后获得了较好的效果,在原代培养时期能较好去除成纤维细胞并通过持续使用该方法可在传二代后获得较高纯度的细胞,纯度能达到80%以上,能够建立细胞系。     

粉纹夜蛾(Trichoplusia ni)培养细胞对家蚕微孢子虫(Nosema bombycis)的吞噬过程观察

观察微孢子虫在非天然宿主细胞系的生物行为过程,有助于探究微孢子虫的侵染特异性机制。选用鳞翅目昆虫粉纹夜蛾(Trichoplusia ni,Tn)细胞系为材料,接种家蚕微孢子虫(Nosema bombycis,Nb)孢子后观察其吞噬过程。结果发现:经氢氧化钾发芽预处理的Nb孢子虽然可以在Tn培养细胞液中发芽,但未见发芽孢子的孢原质进入Tn培养细胞;在接种Nb孢子浓度为2.0×105mL-1以上时,观察到孢子通过内吞作用进入了Tn培养细胞,但未见孢子的增殖;随着时间的延长和进入Tn培养细胞的Nb孢子的增加,可引起Tn培养细胞的超微结构变化,并导致细胞的裂解。以上结果说明,虽未见Nb以发芽形式感染Tn培养细胞,但Nb能够被吞噬进入Tn培养细胞并破坏其结构。     

SR92鸡的OPAY02型标记初步研究

测定了SR92A系鸡初生、28、49、75日龄体重,屠体、半净膛、全净膛重等生产性能,同时用OPAY02随机引物对其DNA进行RAPD扩增,分析OPAY02型标记与鸡群早期增重及75日龄屠宰性能之间的关系。结果表明,片段长度为1 660.8bp和2 326.0bp的两条带所代表的位点,在SR92A系中出现A、B、C三种类型,其中C型母鸡75日龄体重超过A型607.4g,初步表明OPAY02C型标记在SR92A系中可作重要遗传标记。     

Screening of Probiotics Inhibiting E. coli K88 in vitro

The probiotics tested in the experiment were isolated from the intestinal of weaning piglets.The isolated probiotics and E.coli K88 were inoculated into the culture of intestinal porcine epithelial cell-1 (IPEC-1).The activity of lactate dehydrogenase (LDH) in the supernatant was measured after incubating for 2.5 h.At the same time, the probiotics and E.coli K88 were co-cultured in vitro, the number of E.coli K88 was counted and the probiotics which could be resistant to the E.coli K88 were selected 2.5 h later.The results showed that the Lactobacillus casei and Bacillus coagulans could significantly reduce LDH activity (P<0.05), decrease the damage of E.coli K88; Bacillus coagulans could inhibit the growth of E.coli K88.At the same time, Bacillus coagulans could resist high temperature, acid and bile salt.The results showed that Bacillus coagulans strains had great potential as the application of probiotics strains.The methods could be used as a model of screen probiotics which could inhibit the growth of E.coli K88 in vitro.     

多叶苜蓿自交二代农艺性状分析

多叶苜蓿(Medicago sativa L.)的形态特征与苜蓿的产量和品质有直接关系,建立多叶苜蓿自交系进一步研究评估苜蓿多叶性状的遗传特性和规律,为多叶苜蓿新品系的选育提供科学依据。筛选多叶苜蓿自交亲本,进行亲本选配,田间试验观察测量多叶苜蓿自交S2代的多叶率、株高等农艺性状,与亲本材料进行比较分析。发现多叶苜蓿自交S2代农艺性状变异较为丰富,变异系数达19.54%~55.66%,分离出单株多叶率达100%的单株,S2群体生长速率变缓,多叶性状的表达呈多样化。     

川藏黑猪S04系选育研究

选用杜洛克公猪血缘9个（含精液）,母猪50头,组成S04系核心选育群。采用闭锁与开放相结合的选育方法,应用GBS软件进行遗传评估,主选产仔数、日增重与活体背膘厚等性状。经过多个世代的选育,经产母猪产仔数达10．36头,公猪达100kg体重日龄165．67d,活体背膘厚11．16mm,瘦肉率达64．22％,肌内脂肪含量2．88％;主选性状全面达到或超过育种目标,达配套系审定要求,成功培育了一个棕黑色的高瘦肉率专门化父系（S04系）。     

Short‐term effects of cattle‐to‐sheep ratio and stocking rate on patch grazing in sour grassveld

Abstract

The short‐term effects of stocking rate (AUE ha^?1) and cattle‐to‐sheep ratio on the extent and severity of patch grazing and on patch size were evaluated. The extent of patch grazing was proportional to stocking rate, and livestock species had a small but significant effect. Patch size was also a function of stocking rate. Cattle and sheep had similar effects on patch size distribution when stocked at the same number of AUE ha^?1. The severity of grazing within a patch was influenced by both stocking rate and cattle‐to‐sheep ratio. Recommendations that cattle should be grazed together with sheep and that cattle should be in greater proportion (in terms of AUE ha^?1) than sheep, which are aimed at limiting the severity of grazing by sheep, are supported. However, severe grazing within patches is an inevitable consequence of sheep grazing irrespective of the stocking rate or cattle‐to‐sheep ratio applied.     

猕猴桃溃疡病防治研究

对猕猴桃细菌性溃疡病防治方法进行研究,结果表明,冬末春初“划道”控制,病斑,夏委彻底刮除治疗病斑,疗效达80％－90％,DTMz等为此病防治的首选药物。