黑大豆异黄酮浸提的研究

采用石油醚回流技术脱去大豆豆粉中的油脂,将残渣真空烘干后,作为浸提原料.通过比较浸提溶剂、乙醇浓度、浸提温度、液固比、浸提时间和浸提次数等单因素试验结果,结合正交试验设计,得出提取大豆异黄酮的最佳浸提条件:乙醇浓度40%、液固比10∶1、浸提时间3 h、浸提温度90℃.     

鸡的气雾免疫

气雾免疫,省工省力,且对呼吸道有亲嗜性的疫苗特别有效(例如鸡新城疫Ⅱ、Ⅳ系弱毒疫苗和传染性支气管炎弱毒疫苗等),所以近年来采用气雾免疫的鸡群正在逐渐增多。     

不同地区及不同季节的柞蚕寄生蝇RAPD分析

利用RAPD技术对不同地区及不同季节发生危害的柞蚕寄生蝇进行了遗传多态性研究。选取11个随机引物对14份供试材料的基因组DNA进行扩增,共扩增出140条稳定条带,其中128(91.43%)条为多态性条带,表明柞蚕饰腹寄蝇(Blepharipa tibialis)、秋柞蚕寄生蝇(未命名)及栗蚕寄生蝇(未命名)间具有丰富的多态性。各材料间的遗传距离在0.0214~0.5143之间,利用UPGMA法进行聚类分析,14份供试材料可聚为两大类:栗蚕(Dic-tyoplocajaponica)寄生蝇与寄生柞蚕的寄生蝇各聚为一类,其中来自河南省的柞蚕饰腹寄蝇自成一类,来自东北地区的柞蚕饰腹寄蝇与秋柞蚕寄生蝇又各自聚为一类。     

饲粮中添加不同生物制剂对杜寒杂交肉羊生产性能和屠宰性能的影响

本试验旨在比较饲粮添加不同生物制剂对杜寒杂交肉羊生产性能、屠宰性能、组织器官发育及肉品质的影响。采用单因素试验设计,选取平均体重约为32 kg的杜寒杂交F1代肉羊160只,随机分为5组,每组4个重复,每个重复8只。对照组采用基础饲粮,试验组分别添加21 mg/kg莫能菌素、4×109CFU/kg地衣芽孢杆菌、3.2×109CFU/kg酿酒酵母菌、1.1 g/kg复合生物制剂(地衣芽孢杆菌≥6×109CFU/g、酿酒酵母菌≥4×109CFU/g、碱性蛋白酶≥1 000 U/g)。预试期10 d,正试期56 d,每2 d记录1次采食量,每20 d进行1次称重,当复合生物制剂组羊只的平均体重达到约50 kg时,每组选取10只羊进行屠宰,测定其屠宰性能、组织器官重量和肉品质指标。结果表明:1)复合生物制剂组平均日增重、屠宰率显著高于对照组(P0.05);地衣芽孢杆菌组和复合生物制剂组料重比显著低于对照组(P0.05)。2)地衣芽孢杆菌组、酿酒酵母菌组、复合生物制剂组复胃重量均显著高于对照组(P0.05),复合生物制剂组复胃重量占宰前活重比例显著高于对照组和莫能菌素组(P0.05);地衣芽孢杆菌组、酿酒酵母菌组、复合生物制剂组小肠重量及其占宰前活重比例均显著高于对照组、莫能菌素组(P0.05)。3)酿酒酵母菌组肾脏重量占宰前活重比例显著高于对照组(P0.05),其他内脏器官重量占宰前活重比例在各组间无显著差异(P0.05)。4)各组肉品质指标差异不显著(P0.05)。综合得出,从作为饲料添加剂对肉羊生产性能作用效果来看,地衣芽孢杆菌、酿酒酵母菌、复合生物制剂可以替代莫能菌素,地衣芽孢杆菌好于酿酒酵母菌,复合生物制剂最佳。     

Inductive expression of the NOD1 signalling pathway in chickens infected with Salmonella pullorum

1. The aim of this study was to describe the role of Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) receptor signalling in chicken.

2. Tissue-specific expression analysis of NOD1, receptor-interacting serine-threonine kinase 2 (RIPK2), nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase 11 (MAPK11 or p38) by quantitative real-time PCR (qRT-PCR) revealed their wide distribution in various organs and tissues.

3. Salmonella pullorum infection activated NOD1 receptor signalling in vivo and in vitro, resulting in significant induction of downstream signalling molecules RIPK2, NF-κB/p65, MAPK11/p38 and the effector molecules IL-1b and IL-8.

4. Activation of NOD1 by its agonist bacterial γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) in HD11 cells induced the adapter molecular RIPK2 and activated the NF-κB/p65 and MAPK11/p38 pathways, resulting in an increase in IL-8 but not IL-1β. Additionally, inhibition of NOD1 using NOD1-shRNA resulted in downregulation of RIPK2, MAPK11 and IL-8, while NF-κB/p65 and IL-1β were unaltered.

5. These results highlight the important role of NOD1 receptors in eliciting the innate immune response following pathogenic invasion in chicken.     

牛粪不同堆储处理方法微生物数量变化的研究

以牛粪为原料,研究露天堆储、粪池堆储和遮雨堆储三种处理在不同时间段微生物变化规律,为合理有效的处理畜禽粪污提供科学参考.结果各处理在堆储过程中的微生物总数呈山型的变化,其中细菌的数量变化类似于微生物总数的变化;真菌的数量变化呈现一条类似于"W"型曲线;放线菌一直呈下降趋势;微生物总量在15、30、45、60、75 d,处理3与处理1和处理2 差异显著(P<0.05),第30 d处理2与处理1差异显著(P<0.05).     

36个燕麦品种不同部位养分分布格局

分析了天祝高寒草甸区36种不同饲用燕麦(Avena sativa)叶片、籽粒和茎秆中的水分、灰分、粗蛋白、可溶性糖、粗脂肪、磷、钙、粗纤维、中性纤维和酸性纤维含量的分布特征。结果显示:36个燕麦品种叶片中水分、灰分、粗蛋白、可溶性糖和钙含量显著高于籽粒和茎秆(P0.05),平均值分别为6.36%、9.28%、19.82%、13.71%和0.25%;籽粒中粗脂肪和磷含量显著高于叶片和茎秆(P0.05),平均值分别为:4.48%和0.25%;茎秆中粗纤维、中性洗涤纤维和酸性洗涤纤维含量显著高于叶片和种子(P0.05),平均值分别为35.62%、65.18%和56.40%。总之,36种燕麦不同部位的营养价值高低依次为叶片籽粒茎秆。     

Construction and application of a bovine immune-endocrine cDNA microarray

A variety of commercial DNA arrays specific for humans and rodents are widely available; however, microarrays containing well-characterized genes to study pathway-specific gene expression are not as accessible for domestic animals, such as cattle, sheep and pigs. Therefore, a small-scale application-targeted bovine immune-endocrine cDNA array was developed to evaluate genetic pathways involved in the immune-endocrine axis of cattle during periods of altered homeostasis provoked by physiological or environmental stressors, such as infection, vaccination or disease. For this purpose, 167 cDNA sequences corresponding to immune, endocrine and inflammatory response genes were collected and categorized. Positive controls included 5 housekeeping genes (glyceraldehydes-3-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, ribosomal protein L19, beta-actin, beta2-microglobulin) and bovine genomic DNA. Negative controls were a bacterial gene (Rhodococcus equi 17-kDa virulence-associated protein) and a partial sequence of the plasmid pACYC177. In addition, RNA extracted from un-stimulated, as well as superantigen (Staphylococcus aureus enterotoxin-A, S. aureus Cowan Pansorbin Cells) and mitogen-stimulated (LPS, ConA) bovine blood leukocytes was mixed, reverse transcribed and PCR amplified using gene-specific primers. The endocrine-associated genes were amplified from cDNA derived from un-stimulated bovine hypothalamus, pituitary, adrenal and thyroid gland tissues. The array was constructed in 4 repeating grids of 180 duplicated spots by coupling the PCR amplified 213-630 bp gene fragments onto poly-l-lysine coated glass slides. The bovine immune-endocrine arrays were standardized and preliminary gene expression profiles generated using Cy3 and Cy5 labelled cDNA from un-stimulated and ConA (5 microg/ml) stimulated PBMC of 4 healthy Holstein cows (2-4 replicate arrays/cow) in a time course study. Mononuclear cell-derived cytokine and chemokine (IL-2, IL-1alpha, TNFalpha, IFN-gamma, TGFbeta-1, MCP-1, MCP-2 and MIP-3alpha) mRNA exhibited a repeatable and consistently low expression in un-stimulated cells and at least a two-fold increased expression following 6 and 24 h ConA stimulation as compared to 0 h un-stimulated controls. In contrast, expression of antigen presenting molecules, MHC-DR, MHC-DQ and MHC-DY, were consistently at least two-fold lower following 6 and 24 h ConA stimulation. The only endocrine gene with differential expression following ConA stimulation was prolactin. Additionally, due to the high level of genetic homology between ovine, swine and bovine genes, RNA similarly acquired from sheep and pigs was evaluated and similar gene expression patterns were noted. These data demonstrate that this application-targeted array containing a set of well characterized genes can be used to determine the relative gene expression corresponding to immune-endocrine responses of cattle and related species, sheep and pigs.     

Pyruvate kinase is necessary for Brucella abortus full virulence in BALB/c mouse

Brucellosis, caused by a facultative intracellular pathogen Brucella, is one of the most prevalent zoonosis worldwide. Host infection relies on several uncanonical virulence factors. A recent research hotpot is the links between carbon metabolism and bacterial virulence. In this study, we found that a carbon metabolism-related pyruvate kinase (Pyk) encoded by pyk gene (locus tag BAB_RS24320) was associated with Brucella virulence. Determination of bacterial growth curves and resistance to environmental stress factors showed that Pyk plays an important role in B. abortus growth, especially under the conditions of nutrition deprivation, and resistance to oxidative stress. Additionally, cell infection assay showed that Pyk is necessary for B. abortus survival and evading fusion with lysosomes within RAW264.7 cells. Moreover, animal experiments exhibited that the Pyk deletion significantly reduced B. abortus virulence in a mouse infection model. Our results elucidated the role of the Pyk in B. abortus virulence and provided information for further investigation of Brucella virulence associated carbon metabolism.