高地隙喷雾机驱动轮独立式驱动系统转向控制模型研究

车辆转弯时的内外轮速平衡问题是车辆驱动轮独立式驱动技术研究的关键问题。根据车辆转弯时轮速不等的现象,以阿克曼转向原理为基础,对车辆转向模型进行简化,运用运动学分析原理研究了内外轮速与转弯时车速及转向角之间的关系;结合车辆动力学原理建立车辆转弯临界模型,分析了车辆安全转弯临界状态,最后设计了实际应用方案。选择在工作原理和安全控制方面较普通的电控差速器更为简单易行,提高了车辆转向性能、简化了车辆结构、降低了成本。此技术运用到农田高地隙喷雾机上可简化其结构、减轻质量,且在农田行走更灵活方便。     

木醋液对牛粪堆肥理化性质的影响

以牛粪为堆肥原料,研究不同浓度木醋液对牛粪堆肥理化性质的影响。试验采用好氧人工翻堆方式进行。试验共设4个处理,浓度分别为0.2%、0.5%、0.8%的木醋液处理和不添加木醋液的对照处理。通过试验分析温度、含水率、EC、pH值、铵态氮和硝态氮含量等指标随堆肥发酵时间变化的特征。结果表明:各处理组在堆肥发酵过程中pH均在适宜微生物生长的范围内,含水率都保持在55%以上;EC都呈先上升后下降的趋势;添加不同浓度木醋液处理与对照处理相比,都显著提高铵态氮硝态氮含量,从而有效减少堆肥发酵过程中氨气挥发和氮素损失,其中浓度为0.5%木醋液处理效果最好,与对照相比,肥堆升温快,进入高温所需时间短,高温持续时间长,在整个堆肥发酵过程中含水率一直保持在60%~70%之间,发酵结束时物料电导率较低,堆料腐熟快,有利于加快堆肥发酵进程。     

Preparation and Identification of Canine Parvovirus NY Strain VP2 Protein Polyclonal Antibody

This study was aimed to prepare canine parvovirus (CPV) VP2 protein polyclonal antibody.The recombinant expression vector pET28a-CPV-VP2 was constructed and transfromed into E.coli BL21 (DE3),the expression of recombinant proteins was induced by IPTG from which the fusion protein was identified by SDS-PAGE.The target protein was purified and emulsify with adjuvant,the prepared immunogen was inoculated into rabbit by subcutaneous injections to prepare of VP2 protein specific polyclonal antibody.The immuno-activity,titers,neutralization titers of the prepared polyclonal antibody were determined by immunoperoxidase monolayer assay (IPMA).The results showed that the expressed recombinant protein VP2 (rVP2) existed in the form of inclusion body with a molecular weight of 72 ku.The prepared polyclonal antibody titer was 1 600 dilution,the virus titer was 10⁷ TCID₅₀/mL,the neutralizing titer was 1∶2 884.The antibodies showed specific reaction with CPV.In conclusion,rVP2 specific polyclonal antibody showed wonderful immunocompetence,specificity and neutralizing activity,providing foundation for the development of genetic vaccine and clinical therapeutic method.     

The Fluorescent Characterization and Analysis of Two Brucella Attenuated Vaccine Infecting Mouse Macrophagocyte

The experiment was conducted to discuss the difference of binding time of green fluorescent protein B.melitensis M5 (GFP-M5) and B.abortus S19 (GFP-S19) infecting the mouse macrophagocyte (RAW264.7),lysosome,endoplasmic reticulum and golgi body in the initial stage and compare the binding rate of GFP-M5,GFP-S19 with organelle in different timeline,respectively,by confocal laser scanning microscope (CLSM) and flow cytometry.The result showed that GFP-M5 and GFP-S19 were successfully constructed.The intracellular survival ability of Brucella M5,Brucella S19,GFP-M5 and GFP-S19 were not obvisouly affected after infecting RAW264.7.GFP-M5 and GFP-S19 could enter the macrophagocyte in 30 mins,and in 2 h the Brucella could reach lysosome,endoplasmic reticulum and golgi body.In addition,the binding time for two attenuated vaccine did not show differences in 1,2,3 and 4 h.The content of GFP⁺ cell produced by RAW264.7 infected by GFP-M5 and GFP-S19 did not show significant differences (P>0.05).Therefore,the two strains did not have significant differences in the invasion ability in the initial stage of infecting host cell.     

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.     

Comparison of emission estimates for non‐CO2 greenhouse gases from livestock and poultry in Korea from 1990 to 2010

It has often been claimed that non‐carbon dioxide greenhouse gases (NCGGs), such as methane, nitrous oxide and fluorinated greenhouse gases, are significant contributors to climate change. Here we nvestigate emission estimates of methane and nitrous oxide from livestock and poultry production, which is recognized as a major source of those NCGGs, in Korea over the period of 1990 through 2010. Based on the data on livestock and poultry populations, emission estimates of methane and nitrous oxide are first derived based on the Tier 1 approach. Then, the Tier 2 approach is adopted to obtain emission estimates of methane and nitrous oxide from cattle, which are known to be the largest sources of these NCGGs and account for about 70% of emissions from livestock and poultry in Korea. The result indicates that the Tier 2 estimates of methane and nitrous oxide emissions from enteric fermentation and manure management are significantly different from the Tier 1 estimates over the analysis period.     

Isolation and characterization of antimicrobial-resistant Escherichia coli from national horse racetracks and private horse-riding courses in Korea

Limited information is available regarding horse-associated antimicrobial resistant (AR) Escherichia (E.) coli. This study was designed to evaluate the frequency and characterize the pattern of AR E. coli from healthy horse-associated samples. A total of 143 E. coli (4.6%) were isolated from 3,078 samples collected from three national racetracks and 14 private horse-riding courses in Korea. Thirty of the E. coli isolates (21%) showed antimicrobial resistance to at least one antimicrobial agent, and four of the AR E. coli (13.3%) were defined as multi-drug resistance. Most of the AR E. coli harbored AR genes corresponding to their antimicrobial resistance phenotypes. Four of the AR E. coli carried class 1 integrase gene (intI1), a gene associated with multi-drug resistance. Pulsed-field gel electrophoretic analysis showed no genetic relatedness among AR E. coli isolated from different facilities; however, cross-transmissions between horses or horses and environments were detected in two facilities. Although cross-transmission of AR E. coli in horses and their environments was generally low, our study suggests a risk of transmission of AR bacteria between horses and humans. Further studies are needed to evaluate the risk of possible transmission of horse-associated AR bacteria to human communities through horse riders and horse-care workers.     

Apparent ileal digestibility of nutrients and amino acids in soybean meal,fish meal,spray‐dried plasma protein and fermented soybean meal to weaned pigs

This study sought to determine whether fermentation could increase apparent ileal digestibility (AID) of dry matter (DM), nitrogen (N), energy (E) and amino acids (AA) in fermented soybean meal (FSBM) greater than that of soybean meal (SBM) in weaned pigs. Four weaned pigs (10.00 ± 0.30 kg) were surgically equipped with T‐cannulas and randomly followed a 4 × 4 Latin square design of treatments (SBM, FSBM, fish meal and spray‐dried plasma protein). Overall, the fermentation process was able to reduce the amount of anti‐nutritional factors (ANF), including trypsin inhibitors, raffinose and stachyose, in the FSBM diet, which were significantly reduced by 39.4, 92.2, and 92.9%, respectively, as compared to the SBM diet. As a consequence of ANF reduction in FSBM, the AID of DM, N and E as well as AA was significantly greater with FSBM than SBM. Taken all together, the fermentation process improved the nutritional quality of SBM, due to ANF reduction, leading to improvement of digestibility of AA. As such, FSBM can be potentially used as a specialized feed ingredient, especially for young animal diets in an attempt to reduce diet costs.     

Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.