2R,3R-丁二醇和2,3-丁二醇诱导匍匐翦股颖抗病性的比较

以2R,3R-丁二醇和2,3-丁二醇作为诱抗剂,在诱导匍匐翦股颖产生对褐斑病抗性的过程中,重点比较了不同施用方式的诱抗效果,筛选出了诱抗剂的最佳作用方式和浓度。结果表明:匍匐翦股颖接菌后第15d,2R,3R-丁二醇根部注射处理下,100μmol/L的病情指数最低,诱导效果最佳;而与叶面喷施相比,叶面喷施的诱导效果不明显;2,3-丁二醇根部注射处理下,250μmol/L的病情指数最低,诱导效果最佳,而与叶面喷施相比,叶面喷施的诱导效果不明显。结果表明,100μmol/L的2R,3R-丁二醇与250μmol/L的2,3-丁二醇根部注射可有效诱导匍匐翦股颖产生对褐斑病的抗性。     

DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype

A new assay for the detection of swine influenza virus (SIV) was developed with a novel nucleic acid probe——Molecular beacon in this study. The specific primers and molecular beacon probes were designed according to the conserved region of H3 and N2 genes of SIV H3N2 subtype. A digital RT-PCR assay was developed for detection of SIV H3N2 subtype. The results showed that SIV H3N2 subtype could be identified simultaneously on this microarry with high sensitivity and reproducibility,which could reach to 10⁶ dilute viruse. The conclusion was that the digital RT-PCR method could analyze quantitatively the RNA templates.On the identification of H3N2 SIV,the digital RT-PCR method was much more scientific than Real-time quantitative PCR method.     

La（NO3）3对达乌里胡枝子种子活力的影响

用质量浓度为100～1 000 μg/mL的La(NO3)3溶液处理48 h达乌里胡枝子Lespedeza davurica种子来研究其种子活力、种子吸水速率和膜透性。结果表明：质量浓度为200～800 μg/mL的La(NO3)3溶液浸种可促进达乌里胡枝子种子的萌发,提高种子活力,提高种子相对吸水量,增大膜通透性;质量浓度为700～800 μg/mL 的La(NO3)3处理效果最佳,质量浓度大于900 μg/mL则抑制种子萌发。     

AOS-80空气净化机对冬季鸡舍空气的净化作用

在有窗或无窗密闭式鸡舍中,舍内产生的有毒有害气体、微生物和粉尘对鸡的健康与生产性能有很大的影响。本研究选用AOS-80空气净化机,测定其对冬季蛋鸡舍空气的净化效果。选取尺寸、样式和饲养密度等完全相同的2栋蛋鸡舍做对比试验,2栋鸡舍通风采用自动控制装置。在试验鸡舍的屋架上按米字形均匀安装6台净化机,测定2栋鸡舍舍内有毒有害气体和空气细菌总数的浓度。结果表明:2栋鸡舍在适当通风条件下(使舍内温度维持在(15±0.1)℃,对照鸡舍和试验鸡舍的空气细菌总数平均浓度分别为33.3cfu/L和10.6cfu/L(P<0.01),净化机使之降低68.2%;NH3的平均浓度分别为1.71mg/m3和1.22mg/m(3P<0.01),净化机使之降低28.6%;H2S的平均浓度分别为0.670mg/m3和0.643mg/m3(P<0.05),净化机使之降低4.03%;蛋鸡平均周死亡率分别为0.997%和0.607%(P<0.01),净化机使之降低39.0%。本研究结果表明,舍内安装空气净化机能显著降低鸡舍空气细菌总数和有毒有害气体浓度,降低蛋鸡死亡率。     

Immunomodulatory effects of β-glucans on porcine alveolar macrophages and bone marrow haematopoietic cell-derived dendritic cells

The immunopharmacological activities of β-glucans with a backbone of β-1,3/β-1,6-linkages associated with anti-tumor, anti-viral, bacterial and fungal infections have been well documented. Dectin-1, a specific pattern recognition receptor for β-1,3/β-1,6-glucans, is expressed mainly on phagocytes, especially macrophages and dendritic cells (DCs). In this study, the encoding nucleotide for the carbohydrate-recognition domain (CRD) of porcine dectin-1 was sequenced for the first time, and the immunomodulatory functions of a synthetic particulate β-glucan (p-β-glucan) were examined. Results showed that p-β-glucan significantly enhanced cell activity and phagocytosis in porcine alveolar macrophages (AMs), immature DCs (imDCs) and mature DCs (mDCs), in a similar way to zymosan. Zymosan enhanced dectin-1/TLR2/TLR4 expression and TNF-α/IL-10 production in all of three types of cell, whereas p-β-glucan increased dectin-1/TLR4 and TNF-α/IL-12 production in AMs but inhibited IL-10 in mDCs. These results indicate that the complex collaborating interactions between dectin-1 and TLRs in the recognition of β-1,3/β-1,6-glucans with different structural features may direct different cellular responses.     

Molecular Characterization of Equine Rotavirus Group A Detected in Argentinean Foals During 2009–2014

Equine rotavirus group A (RVA) has been detected in several countries worldwide since its first detection in 1975. Currently, equine RVA is considered the major cause of dehydrating diarrhea in foals younger than 3 months, and the frequency of detection in clinical cases varies from 20% to 77%. The genotypes of epidemiologic relevance found in horses are G3P[12] and G14P[12]. In a survey conducted in Argentina from 1992 to 2008, equine RVA was detected in 21% and 39% of the fecal samples and outbreaks, respectively. Genotype distribution was 51% G3P[12] and 33% G14P[12]. In continuation with the surveillance, the aim of the present study was to characterize the equine RVA detected in Thoroughbred foals in Argentina from 2009 to 2014. A total of 436 stool samples (corresponding to 177 single diarrhea cases or outbreaks) were analyzed. Equine RVA was detected in 31% (135 of 436) of the samples, which corresponded to 42% (74 of 177) of outbreaks. From the positive cases, 42% (57 of 135) were genotyped. Of this, 63% were G3 (36 of 57) and 37% (21 of 57) were G14 genotype. Considering the whole data (1992–2014), equine RVA was detected in 25% (300 of 1,207) of the stool samples and 41% (119 of 293) of the diarrhea outbreaks. The results of this study also show a cyclic pattern of the G3 and G14 prevalence in the horse population with a change in G3:G14 frequencies from year to year. Furthermore, clustering in the phylogenetic tree suggests evolutionary and geographic relationships between the Argentinean strains compared with the strain circulating worldwide.     

H3亚型猪流感病毒荧光定量PCR检测方法的建立

通过RT-PCR方法克隆了H3亚型猪流感病毒HA基因一段靶序列,构建重组质粒作为标准阳性模板.根据GenBank中的H3亚型猪流感病毒HA基因保守序列设计了用于FQ-PCR的1对引物和1条TaqMan探针.通过条件优化,以10倍系列稀释的质粒为标准品进行荧光定量PCR扩增,并制作标准曲线,建立了检测H3亚型猪流感的荧光定量PCR方法.结果表明,该方法检测灵敏度可达1.0×100拷贝/μL,线性范围为109～100,达10个数量级;对起始浓度为1.0×109、1.0×108、1.0×107拷贝/μL的标准品的最终实际测得值(Ct)分别为13.68,18.21和20.57;变异系数分别为0.31%、0.17%和0.12%,均小于5%,说明此方法具有良好的准确性和重现性.对阳性组织病料的检测表明,该方法的检测灵敏度高出常规PCR,与套式PCR具有相近的灵敏度.     

日粮中添加戊酸对奶牛体外发酵部分指标的影响

本试验以装有瘤胃瘘管的3头荷斯坦奶牛为试验动物,利用体外批次培养试验法.研究了在奶牛三种不同品质的粗饲料日粮中分别添加不同浓度的戊酸(1.25、2.50、5.00、10.00mg/g)对其体外发酵pH、氨氮(NH3-N)浓度和气体产量的影响.结果表明:(1)培养液pH值均随着培养时间的延长呈逐渐下降的趋势,但各试验组和对照组均值差异不显著(P>0.05).(2)培养液NH3-N浓度的变化趋势基本一致.随着培养时间的延长,培养液NH3-N浓度在3～6 h降低,9～24 h又逐渐升高.(3)培养液气体产量的变化均随培养时间的延长呈逐渐的上升趋势.     

2型猪圆环病毒感染对小鼠组织中Caspase3、Bak及Bcl-2基因表达的影响

Caspase3、Bcl-2、Bak是细胞凋亡过程中3种重要的调节蛋白,为研究感染猪圆环2型病毒(PCV2)后体内细胞的凋亡情况,本实验以BALB/c小鼠为实验模型,建立了SYBR Green Ⅰ实时定量PCR检测细胞凋亡的方法.结果表明实验组Caspase3的表达在各时间段里与对照组相比均呈上升趋势,提示了PCV2感染可以上调Caspase3水平,从而增加被病毒感染组织的细胞凋亡,而小鼠心、脑、脾等组织Bak基因表达明显上调,Bcl-2基因表达水平也伴随Bak基因而升高.这些结果揭示了PCV2感染BALB/c小鼠后,相关细胞凋亡基因在体内的作用与机制,同时为今后细胞凋亡的检测提供了新的方法.