鸭疫里氏杆菌吉林分离株camp基因的克隆、表达及间接ELISA检测方法的建立

为建立鸭疫里氏杆菌(RA)的血清学快速检测方法,本研究根据GenBank中登录的camp基因序列设计引物,以吉林RA分离株JL-1的基因组DNA为模板,通过PCR扩增camp基因,并构建pET-camp重组表达质粒,转化至E.coli BL21 (DE3)中诱导表达出分子量约为37 ku重组蛋白,westemblot分析结果表明该蛋白能够与血清1、2、10、11和17型RA全菌体阳性血清发生特异性反应.以纯化的重组蛋白为包被抗原建立间接ELISA方法,结果显示其具有良好的特异性和重复性,与RA超声裂解抗原ELISA方法的符合率达到77.5％.研究结果进一步验证了Camp蛋白具有良好免疫原性,并且为RA多种血清型共同抗原,在RA的检测及亚单位疫苗开发中具有良好的应用价值.     

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.     

盐胁迫对5份无芒雀麦苗期生长和生理生化的影响及综合性评价

为了明确不同无芒雀麦苗期耐盐能力的强弱,本研究选择5份不同株型的无芒雀麦作为供试材料。根据新疆土壤盐碱成分设置中性盐(M盐)：NaCl∶Na₂SO₄∶Na₂CO₃=1∶4∶0和碱性盐(A盐)：NaCl∶Na₂SO₄∶Na₂CO₃=1∶1∶8两种处理,于三叶期每盆一次性浇灌2 L电导率(EC)为20 ms·cm^-1的盐处理液,CK浇灌等量自来水。处理30 d后测定生长指标、叶绿素荧光特性参数、丙二醛(MDA)和渗透调节物质含量,并通过相关性分析、主成分分析、隶属函数分析对13个指标进行分析评价。结果表明：除F_v/F_m和F_v/F_o外,盐胁迫显著影响其余11个指标,且5份材料间差异显著(P<0.05);5份无芒雀麦的株高、茎粗、分蘖数、地上生物量、根冠比、MDA和脯氨酸含量在不同盐胁迫下表现出不同的适应变...     

鸭疫里氏杆菌免疫原性研究Ⅰ.荚膜的形态学观察及荚膜物质的提取

本研究应用电镜对Ⅰ型鸭疫里氏杆菌的荚膜进行了初步观察,在电镜下,负染的菌体周围呈一电子透明层;未经抗体处理的细菌超薄切片,荚膜在菌体周围呈松散的絮状结构,而应用同源抗体处理后,荚膜则呈一均匀层。同时在超薄切片中还看到细菌的空泡和外膜结构。采用十六烷基三甲基溴化铵（ＣＴＡＢ）成功地提取了培养物上清液中的荚膜成分,经过测定比较,加入５ｍＭＣＴＡＢ时效果最佳。     

Green tea polyphenols supplementation alters immunometabolism and oxidative stress in dairy cows with hyperketonemia

Peripartal cows often experience negative energy balance, and are therefore prone to suffering from metabolic diseases such as hyperketonemia, which causes financial losses in dairy farms. This study aimed to investigate the effect of green tea polyphenol (GTP) supplementation during the periparturient period on production performance, oxidative stress and immunometabolism in dairy cows with hyperketonemia. One hundred Holstein cows were assigned to GTP (0.2 g/kg DM; n = 50) or control (without GTP; n = 50) group based on body weight, previous milk yield, and parity on d 15 before expected parturition. Subsequently, 10 cows with hyperketonemia were selected from each group, according to blood β-hydroxybutyric acid (BHBA) concentration between 1.2 and 2.9 mmol/L from 2 to 3 d postpartum. All cows were fed a close-up diet and a lactation diet with or without GTP supply from 15 d prepartum until 30 d postpartum. Milk and blood samples were obtained from 20 cows selected with hyperketonemia on 10, 20, and 30 d postpartum. Compared with control cows, greater milk yield and lower somatic cell count were observed in GTP cows. The GTP group had lower concentrations of BHBA, free fatty acids, cholesterol, triglyceride, reactive oxygen species, malondialdehyde, and hydrogen peroxide, greater concentrations of glucose, lower activities of aspartate aminotransferase, alanine aminotransferase, and glutamyl transpeptidase, alongside greater activities of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity. Additionally, GTP supplementation up-regulated concentrations of interleukin-6 and interleukin-10, but down-regulated concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-2, interleukin-8, and interferon-γ in plasma. Greater concentrations of plasma immunoglobulin G were also detected in the GTP group. Overall, the data suggested that GTP supplementation from 15 d prepartum to 30 d postpartum improved the milk yield and health status in cows with hyperketonemia during early lactation.     

鸡lncRNA-MSTRG.15568.9及其预测靶基因的表达

试验旨在了解在鸡睾丸中高表达的1个长链非编码RNA （long non-coding RNA,lncRNA）及其预测靶基因的时空表达规律,研究二者在鸡弱精子症中的调控作用。根据弱精子症和正常北京油鸡公鸡睾丸转录组测序筛选到的1个高表达的lncRNA （MSTRG.15568.9）,采用顺式（cis）作用模式预测其潜在靶基因SPAG4（sperm-associated antigen 4）,进一步采用实时荧光定量PCR方法进行表达量分析。分别选择3只0、5、20、30、45、60周龄正常北京油鸡公鸡,检测MSTRG.15568.9与SPAG4基因在不同周龄公鸡睾丸中的表达量差异;选择30周龄3只正常公鸡,采集睾丸、肝脏和脾脏等8个部位组织样品,检测MSTRG.15568.9与SPAG4基因在不同组织间的表达规律;选择45周龄弱精子症公鸡和正常公鸡各3只,对比MSTRG.15568.9与SPAG4基因在睾丸的表达量差异。结果显示,MSTRG.15568.9与SPAG4存在明显的时空表达差异,且二者表达趋势基本一致。在不同周龄的鸡睾丸组织中,MSTRG.15568.9和SPAG4的表达趋势相近,MSTRG.15568.9在20周龄的表达量显著高于0、5、30、45、60周龄（P<0.05）,0和5周龄表达量显著低于20、30、45和60周龄（P<0.05）;SPAG4在45周龄表达量最高,其次是20周龄（P<0.05）。MSTRG.15568.9和SPAG4在睾丸和肝脏中的表达量均显著高于脾脏、肾脏等组织（P<0.05）;在正常睾丸组织中的表达量均显著高于弱精子症睾丸组织（P<0.05）。综上所述,MSTRG.15568.9与SPAG4基因具有较明显的组织表达特异性,且MSTRG.15568.9可能调控SPAG4基因的表达,参与精子发生与精子活力调控;但其具体作用机制需要进一步探索。本研究可为鉴定与鸡弱精子症调节机制相关的功能基因提供参考。     

Pharmacokinetics of sanguinarine,chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration

Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit®. However, little information is available on the pharmacokinetics of Sangrovit® in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit® and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half‐life (t_1/2) values of 1.05 ± 0.18 hr and 0.83 ± 0.10 hr for SA and DHSA, respectively. The maximum concentration (C_max) of DHSA (2.49 ± 1.4 μg/L) was higher that of SA (1.89 ± 0.8 μg/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 ± 5.4 and 6.08 ± 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 ± 0.63 (L·h^?1·kg^?1) with a t_1/2 of 0.34 ± 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 ± 1.05 and 0.52 ± 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit® had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit® after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.     

星星草对碱化土壤物理性质的影响

研究不同生长年限的星星草对碱化土壤及对应碱斑土壤的机械组成、土壤孔隙度、容重和含水量的影响。结果表明,星星草生长1、2和3年后,粘粒和细砂所占比例减小,而粗粉和细粉所占比例增大。土壤总孔隙度、毛管孔隙度和非毛管孔隙度随着生长年数的增加呈不同程度的增加,而土壤容重则越来越小,>0.25mm的水稳定性团聚体随之增加。星星草地土壤含水量和田间持水量均高于碱斑土壤。星星草改善了碱化土壤结构、通透性、保水性和土壤物理环境。     

返滴定法测定蜂胶中游离酸含量

建立测定蜂胶品中游离酸含量的方法.采用弱碱溶解蜂胶再酸化萃取的方法纯化样品,用返滴法测定,以水杨酸作为对照品检测样品中游离酸含量.结果测定了5个不同产地的蜂胶样品,游离酸含量在1.89～6.54%范围内,回收率为99.30%.本方法操作简单,重现性好,有助于蜂胶质量的鉴别.     

玉米副产物的饲用价值

本文主要介绍了玉米加工的副产物的种类、数量及常见几种主要玉米副产物的营养价值,并进一步阐述了其饲用畜禽的饲用价值。