嗜吞噬细胞无浆体SYBR GreenⅠ荧光定量PCR检测方法的建立及应用

为建立荧光定量PCR方法检测嗜吞噬细胞无浆体(Anaplasma phagocytophilum),针对GenBank A.phagocytophilum 16S rRNA基因保守序列(JN558815),设计1对引物1-25F/183-205R。以已知引物EE1/EE2和该引物进行巢式PCR扩增出预期大小片段(205bp),构建含有16S rRNA基因的重组质粒,以质粒为模板构建了标准曲线。以1-25F/183-205R为荧光定量PCR引物,建立A.phagocytophilum荧光定量PCR检测方法,并进行特异性、敏感性、重复性试验和临床样本检测。结果表明,该方法在6.4×10~3~6.4×10~8 copies/μL相关系数为0.99,显示出良好的线性关系;所建方法能特异地检出A.phagocytophilum,对绵羊无浆体、牛无浆体和环形泰勒虫基因组DNA和灭菌双蒸水均无交叉反应;可检测到6.4×10~2 copies/μL的标准质粒DNA,比常规PCR敏感性高100倍;批内及批间变异系数均低于2.0%,具有较高的重复性和稳定性;利用该方法对30份羊血液临床样品检出率比巢式PCR高16.67%。该研究为A.phagocytophilum临床检测和定量分析病原感染程度奠定理论基础。     

猪瘟病毒NASBA-RT-LAMP快速检测方法的建立

本研究基于核酸序列依赖性扩增技术(NASBA)和反转录环介导等温扩增技术(RT-LAMP),建立了一种针对猪瘟病毒的,快速、灵敏、特异的两步法检测技术。该方法的检测灵敏度与反转录巢式PCR(RT-NestPCR)相当,最低能检测出46 pg/μL的病毒RNA。该方法的建立为猪瘟病毒的快速诊断提供了新思路。     

羊种布鲁氏菌疫苗株PCR-RFLP鉴定方法的建立

为建立一种羊种布鲁氏菌Rev.1疫苗株PCR-RFLP鉴定方法,利用羊种布鲁氏菌Rev.1具有链霉素抗性的特性,选取与链霉素抗性相关编码核糖体蛋白S12的rps L基因为模板设计引物,经PCR扩增后,将产物进行Nci I酶切,发现羊种布鲁氏菌疫苗株Rev.1出现约500 bp条带,而不具有链霉素抗性的羊种布鲁氏菌株16M则出现384 bp条带。结果表明,PCR-RFLP方法能够用于羊种布鲁氏菌Rev.1疫苗株的鉴定,这为今后区分羊种布鲁氏菌疫苗株Rev.1免疫与野株感染提供了基础。     

不同成熟阶段及品种对全株小麦营养成分和瘤胃降解特性的影响北大核心CSCD

本研究旨在通过研究不同成熟阶段及品种对全株小麦营养成分和瘤胃降解特性的影响,为全株小麦应用于奶牛饲粮提供数据参考。试验采用山农22026(SN22026)和山农82567(SN82567)2个品种的小麦,样品采集期为2015年3月下旬至6月中旬。试验测定了不同成熟阶段全株小麦干物质(DM)、粗蛋白质(CP)、淀粉、粗灰分、有机物、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)、酸性洗涤木质素(ADL)含量及饲料相对饲喂价值,并用3头体重约500 kg的装有瘤胃瘘管荷斯坦奶牛测定了小麦瘤胃降解率。结果表明:1)蜡熟期全株小麦的DM含量显著高于其他各期(P<0.05);全株小麦的CP含量在成熟过程中逐渐减少,开花期后CP含量保持稳定;随着小麦的成熟,全株小麦的NDF、ADF和ADL含量呈先增加后减少的趋势。2)全株小麦单位DM的总能在成熟过程中呈下降趋势,泌乳净能呈先下降后增加的趋势。3)全株小麦DM、CP和NDF的有效降解率在拔节期最高,抽穗期和开花期显著降低(P<0.05),乳熟期和蜡熟期又显著升高(P<0.05)。4)全株小麦的瘤胃降解蛋白质(RDP)含量在拔节期最高,显著高于其他各期(P<0.05);抽穗期显著降低(P<0.05),开花期降至最低(P<0.05),开花期后逐渐升高(P>0.10);而瘤胃非降解蛋白质含量在小麦成熟过程中逐渐下降,蜡熟期最低,显著低于其他各期(P<0.05)。由此可见,本试验2个品系全株小麦相比,仅在几个指标中有显著差异。每个成熟阶段的全株小麦饲养价值都很高,但不同成熟阶段全株小麦的营养价值、产量和瘤胃降解特性存在显著的差异。     

‘银森’女贞扦插繁殖

<正>‘银森’女贞(Ligustrum japonicum‘Jack frost’)为木犀科女贞属常绿丛状灌木,又名‘银霜’,是日本大叶女贞的一个栽培品种。树高可达2~3m,枝干青色,有白色斑点。单叶对生,叶片卵形,先端尖,叶革质,具蜡质,叶片边缘银白色,质厚光亮。4~5月,当年生枝先端发出圆锥花序,花朵白色,花冠筒状漏斗形,呈4深裂。核果,椭圆形,成熟时果实呈黑紫色。‘银森’女贞喜光,耐旱性强,侧根发达,萌芽力强,在酸性、中性和微碱性土中均可正常生长。‘银森’女贞分布于日本关东以西,本州、四国、九州以及中国台湾。2001年由浙江森禾种     

Research Progress on Bacterial Resistance in Environment

Antibiotic resistance genes have become a recognized environmental pollutant, threatening the ecological safety and human health. The application of antibiotics in the clinical and animals breeding, making the environmental microorganisms living with the impact of the residue of antibiotics and elements of resistance genetic and leading to antibiotic resistant bacteria to gain a competitive advantage and destroyed the stability of ecosystem. In this paper, we expounded the concept of resistance gene transmission by the view of macro environment. Through analyzing the mechanism of resistance development,environmental pollution caused by drug resistance and the impact of environmental microorganism for drug resistance, we clarified the key role of the environment in the development of the characteristics of bacterial resistance and analysis environmental resistance.Such as the ecological diversity of flora, the types and the spread of resistant bacteria, the residues of antibiotics and the transmission of the resistance genes.     

优质常规水稻品种美香占2号高产栽培技术

分析了美香占2号品种特性,介绍其高产栽培技术,包括选种、育秧、移栽、肥水管理以及病虫草害防治等方面内容,以对提高水稻产量起到一定的参考作用。     

表达PCV2 CAP蛋白和猪IL-18重组PRV PGO18株的生物学特性

为了探讨表达猪圆环病毒2型(PCV2)CAP蛋白和猪IL-18的重组猪伪狂犬病病毒PGO18作为疫苗候选株的潜在价值,对其在ST细胞上的培养特性,ST、VERO、IBRS-2、PK-15和IPEC细胞上增殖情况,生长特性,遗传稳定性,对小鼠安全性及其理化特性等生物学特性进行了研究。结果表明,重组病毒PGO18株与亲本株猪伪狂犬病病毒HB98具有相似的培养特性和生长特性,在ST、VERO和IBRS-2细胞上的增殖滴度TCID50与亲本株(107.75/0.1mL)相当,遗传稳定性良好,安全性试验表明对小鼠是安全的;理化特性试验说明该病毒具有猪伪狂犬病病毒的通性。     

一起羊小反刍兽疫的诊断及其病毒分离、鉴定和全基因组序列分析

2014年8月,河北涿鹿某养殖场羊群发生不明原因的疫病,临床症状表现为腹泻,咳嗽,呼吸困难,口腔有溃疡,流鼻涕,发热,流泪。应用RT-PCR技术从病羊体内扩增出了小反刍兽疫病毒(PPRV)的基因片段。序列分析发现,China/BJ/2014与China/XJYL/2013的同源性最高,为99.7%,并将该毒株命名为China/BJ/2014。遗传进化树分析表明,China/BJ/2014属于PPRVⅣ系,与China/XJYL/2013、CH/HNZK/2014和CH/GDDG/2014亲缘关系最近。全基因组序列分析发现,与另外3株序列相比,China/BJ/2014存在15处氨基酸替换。本研究确定了此次疫病为小反刍兽疫,且该毒株为国内流行株,全基因组序列分析为进一步研究PPRV进化和防控提供了重要信息。     

Effect of Different Concentrate-roughage Ratios on Rumen Degradability of Cassava Stems and Leaves Total Mixed Ration in Goat

The rumen degradations of TMR were studied in this experiment,in which cassava stems and leaves were used as roughage and formulated with concentrate in different proportions.The purpose of this study was to formulate the available TMR for finishing goat,and provide a theoretical basis for the applications of cassava stems and leaves in goat production.Six Hainan Black goats with permanent rumen fistulas were selected,the rumen degradabilities of DM,CP,NDF and ADF of TMR with 6 different concentrate to forage ratio（2:8,3:7,4:6,5:5,6:4 and 7:3）were tested using nylon bag method.The results showed that,the DM effective degradabilities at the ratio of 4:6,5:5,6:4,7:3 were significantly higher than that of 2:8 and 3:7（P<0.05).CP effective degradabilities at the ratio of 2:8,3:7,4:6 were significantly higher than that of 5:5,6:4 and 7:3（P<0.05).The NDF effective degradation at the ratio of 5:5 was significantly higher than that in the other groups（P<0.05).The ADF degradation at the ratio of 3:7 was significantly higher than that of 4:6,5:5, 6:4 and 7:3（P<0.05).The results indicated that,the nutrient degradation in rumen were higher when the concentrate to forage（cassava stems and leaves）ratio in TMR were 3:7,4:6 and 5:5,and the recommended ratio range were 4:6 to 5:5 in production practice.