兔巴氏杆菌和波氏杆菌多重PCR检测方法的建立

参考GenBank中兔巴氏杆菌16SrRNA和波氏杆菌的fim2的基因序列,应用Premier 5.0软件在二者高度保守区设计了2对引物,建立了适合巴氏杆菌和波氏杆菌快速检测的多重PCR检测方法。以该方法对巴氏杆菌和波氏杆菌参考菌株进行PCR扩增,分别能从各自的基因组中扩增出与试验设计相符的644bp和425bp的特异性DNA片段。将扩增所得的DNA片段进行克隆测序,测序结果表明分别为巴氏杆菌16SrRNA和波氏杆菌fim2基因序列。该方法对波氏杆菌的检测下限为4×102 CFU,对巴氏杆菌的检测下限为6×101 CFU。对兔源性沙门菌、葡萄球菌、大肠杆菌、魏氏梭菌扩增结果为阴性。表明所建立的RT-PCR检测技术具有特异、快速和敏感的特点,可用于鉴别诊断兔巴氏杆菌和波氏杆菌以及2者混合感染。     

H5亚型禽流感病毒一步法复合RT-PCR检测方法的建立

针对近年来H5亚型禽流感(AI)频发的严峻形势,本研究通过分析GenBank中AI的221个HA基因和M基因序列,在保守区内用Oligo4.0软件设计合成2对引物,建立了能同时鉴别AIV型与亚型的一步法复合RT-PCR,其扩增的目的片段大小分别为372、229 bp。通过对AIV尿囊液和棉拭子浸出液进行不同稀释倍数检测,结果病毒在尿囊液最低检出量为10-4稀释。用病毒分离和该方法同时检测不同脏器、口咽及泄殖腔棉拭子样品,结果该方法检测灵敏度比病毒分离低10~100倍。用该方法检测H1~H15亚型AIV和鸡新城疫病毒等其他14种禽病毒,结果所有AIV均有M基因229 bp的目的条带,且仅H5亚型AIV有372 bp的特异性条带,而其他14种禽病毒无任何目的条带。     

低聚木糖与抗生素组合应用对肉用仔鸡生产性能的影响

低聚木糖由2～8个木糖以β-1,4糖苷键连接而成,有效成分木二糖、木三糖、木四糖、木五糖等,以木二糖和木三糖为主,是一种功能性寡聚糖类益生元.低聚木糖作为饲料添加剂具有添加量少、成本低、效益高等优点.目前在畜牧生产上对果寡糖、甘露寡糖等低聚糖类添加剂已有大量的研究报道,而对低聚木糖与抗生素组合应用或替代抗生素的相关研究报道较少.本试验旨在通过研究低聚木糖与抗生素组合添加对肉鸡生产性能的影响,探讨其配伍效应,为合理应用低聚糖类饲料添加剂,降低抗生素在饲料中的用量提供参考依据.……     

烤烟生产打叶组合配方计算机辅助系统构架研究

本文分析了当前烤烟生产与打叶复烤之间存在的主要问题,从烟叶的生产、收购、仓储、品质及叶组配方等5个方面进行构架建设,提出一个以烟叶主要化学成分作为品质依据进行打叶复烤组合配方的计算机辅助系统解决方案.     

放牧强度对羊草草原的影响

研究放牧强度对羊草草原植物群落组成、数量特征、生物量及土壤特性等的影响.结果表明,随着放牧强度的增加,羊草草原的植被盖度和生物量随之降低,优势羊草群落将逐渐被盐生植物所替代,群落结构趋于简化,物种向旱生化和盐生化演替.同时土壤容重、含水量、有机质和氮、磷、钾、钙、镁等营养元素含量亦随着放牧强度的增加而逐渐降低.春季过度放牧对土壤表层的水分、有机质和钙、镁等元素造成的损失最为严重.因此,制定科学的草原管理措施,特别是春季限制放牧等措施对于维持草原的可持续发展具有重要意义.     

土壤速效磷浓度对白三叶构型及能量分配的影响

土壤速效磷浓度对白三叶地上部的垂直分布影响显著。结果表明,白三叶构件密度、分枝密度、分枝角度及总能量均与土壤速效磷浓度呈极显著正相关,节间长度则与之呈极显著负相关。叶片和匍匐茎的能量与土壤速效磷浓度呈正比。花序的能量则随着有效磷浓度的增加而减少。茎能量分配随着有效磷浓度的增加呈铃形曲线型变化。     

蛋鸡种蛋蛋重对孵化率和雏鸡生长发育的影响

将豫州褐壳蛋的种蛋按重量大小分成6组，在相同的条件下进行孵化和饲养试验。结果表明，蛋重在53－56克的种蛋，其受精蛋孵化率最高；不同的蛋重对雏鸡的初生重以及雏鸡在1－3周龄的体重影响较大，蛋重与初生重（γ=0．9899）、1周龄体重（γ＝0．9528）的相关极显著；蛋重与2周龄体重（γ＝0．9032）、3周龄体重（γ＝0．9040）相关显著，与3周龄以后雏鸡体重相关不显著。     

人工模拟高温环境下白三叶草相对生理指标测定

研究了高温胁迫条件下白三叶叶片相对含水量、膜透性、脯氨酸含量和叶绿素含量的变化规律。结果表明:25~35℃处理0.5 h后,白三叶的膜透性、脯氨酸含量逐渐上升,相对含水量、叶绿素含量逐渐下降,但幅度不大;45℃以上高温处理后,以上生理指标均有显著性大幅度的变化。与CK比较,温度每升高5℃,白三叶细胞质膜透性、脯氨酸含量平均增加50.62%、59.89%,而相对含水量、叶绿素含量分别下降34.22%、4.61%。在实验条件下,白三叶能适应35℃的高温,40~45℃为其忍耐高温的临界温度范围,50℃为致死温度上限。     

一株饲用枯草芽孢杆菌CCAM080032固态发酵工艺的研究

为了提高一株饲用枯草芽孢杆菌CCAM 080032的芽孢产量,研究通过单因素试验及正交试验方法,确定了该枯草芽孢杆菌的最优培养基为:稻草粉:麸皮2:8、葡萄糖50 g、胰蛋白胨20 g、NH4NO3 15 g、KH2PO4 10 g、H2O 1 500 g,37℃培养48 h,芽孢产量达到7.90×1010 CFU/g。     

The 16MΔvjbR as an ideal live attenuated vaccine candidate for differentiation between Brucella vaccination and infection

Brucellosis brings great economic burdens for developing countries. Live attenuated vaccines are the most efficient means for prevention and control of animal Brucellosis. However, the difficulties of differentiating of infection from vaccine immunization, which is essential for eradication programs, limit their applications. Therefore, the development of a vaccine that could differentiate infection from immunization will overcome the limitations and get extensive application. VjbR is a quorum sensing regulator involving in Brucella's intracellular survival. The vjbR∷Tn5 mutants have been proven effective against wild type strain challenge, implying its possibility of use in vaccine candidate development. To further evaluate this candidate gene, in the present study, the antigenicity of purified recombinant VjbR protein was analyzed. Antibodies to Brucella melitensis VjbR could be detected in sera from patients and animals with brucellosis but not in control ones, implying the potential use of this protein as a diagnostic antigen. Then a vjbR mutant of B. melitensis 16M was constructed by replacing the vjbR with kanamycin gene. The mutant showed reduced survival in macrophage and mice. Vaccination of BALB/c mice with 16MΔvjbR conferred significant protective immunity against B. melitensis strain 16M challenges, being equivalent to which induced by the license vaccine Rev.1. The vjbR deletion mutant elicited an anti-Brucella-specific immunoglobulin G response and induced the secretion of gamma interferon and interleukin-10. The most importance is that, the use of vjbR mutants as vaccines in association with diagnostic tests based on the VjbR antigen would allow the serological differentiation between infected and vaccinated animals. These results suggest that 16MΔvjbR is an ideal live attenuated vaccine candidate against B. melitensis and deserves further evaluation for vaccine development.