Characterization of an intravenous lipopolysaccharide inflammation model in calves with respect to the acute-phase response

Our objective was to develop a lipopolysaccharide (LPS) inflammation model in calves to evaluate the acute-phase response with respect to the release of pro-inflammatory cytokines and acute-phase proteins, fever development and sickness behaviour. Fourteen 4-week-old male Holstein Friesian calves were included and randomly assigned to a negative control group (n = 3) and an LPS-challenged group (n = 11). The latter received an intravenous bolus injection of 0.5 μg of LPS/kg body weight. Blood collection and clinical scoring were performed at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 12, 18, 24, 28, 32, 48, 54 and 72 h post LPS administration (p.a.). In the LPS group, the following clinical signs were observed successively: tachypnoea (on average 18 min p.a.), decubitus (29 min p.a.), general depression (1.75 h p.a.), fever (5 h p.a.) and tachycardia (5 h p.a.). Subsequent to the recovery from respiratory distress, general depression was prominent, which deteriorated when fever increased. One animal did not survive LPS administration, whereas the other animals recovered on average within 6.1 h p.a. Moreover, the challenge significantly increased plasma concentrations of tumour necrosis factor-α, interleukin 6, serum amyloid A and haptoglobin, with peaking levels at 1, 3.5, 24 and 18 h p.a., respectively. The present LPS model was practical and reproducible, caused obvious clinical signs related to endotoxemia and a marked change in the studied inflammatory mediators, making it a suitable model to study the immunomodulatory properties of drugs in future research.     

Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca²⁺ mobilization of the cells was evaluated using the Ca²⁺ indicator Fluo3 to monitor Ca²⁺ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca²⁺ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.     

Hypoxia inducible factor 1α expression and effects of its inhibitors in canine lymphoma

Hypoxic conditions in various cancers are believed to relate with their malignancy, and hypoxia inducible factor-1α (HIF-1α) has been shown to be a major regulator of the response to low oxygen. In this study, we examined HIF-1α expression in canine lymphoma using cell lines and clinical samples and found that these cells expressed HIF-1α. Moreover, the HIF-1α inhibitors, echinomycin, YC-1 and 2-methoxyestradiol, suppressed the proliferation of canine lymphoma cell lines. In a xenograft model using NOD/scid mice, echinomycin treatment resulted in a dose-dependent regression of the tumor. Our results suggest that HIF-1α contributes to the proliferation and/or survival of canine lymphoma cells. Therefore, HIF-1α inhibitors may be potential agents to treat canine lymphoma.     

氟吡呋喃酮与8种杀虫剂复配对温室白粉虱的联合毒力

为了筛选出对温室白粉虱高效的农药复配组合,本试验选择9种杀虫单剂对田间采集的温室白粉虱进行了毒力测定,并利用共毒因子和共毒系数对氟吡呋喃酮与8种杀虫剂复配组合的联合毒力进行评价,同时对筛选出的药剂进行田间药效试验。结果表明：杀虫单剂毒力测定中,LC₅₀最低的为鱼藤酮,最高的为螺虫乙酯,48 h LC₅₀分别为0.43μg/mL和78.59μg/mL;毒力大小顺序为鱼藤酮>氟啶虫胺腈>除虫菊素>苦参·印楝素>d-柠檬烯>氟吡呋喃酮>啶虫脒>高效氯氰菊酯>螺虫乙酯。共毒系数大于120的复配组合为氟吡呋喃酮与d-柠檬烯5∶1、氟吡呋喃酮与苦参·印楝素1∶2和1∶5,共毒系数分别为268.31、247.80和241.46,LC₅₀分别为5.76、4.51μg/mL和4.34μg/mL。氟吡呋喃酮与d-柠檬烯5∶1复配增效作用明显,为最佳配比。田间药效试验表明,10 d后,17%氟吡呋喃酮可溶液剂与1%苦参·印楝素乳油、5%d-柠檬烯可溶液剂复配对温室白粉虱的防效达到83.5%～83...     

葡萄VvSUC12基因启动子的克隆及上游调控因子鉴定

枝干病害葡萄溃疡病近年来严重限制了葡萄产业发展。研究表明葡萄蔗糖转运蛋白参与寄主植物和病原菌的互作过程。为解析蔗糖转运蛋白VvSUC12在葡萄免疫反应过程中的功能,本研究克隆了VvSUC12基因翻译起始位点上游1 500 bp的启动子区,通过生物信息学分析发现该区域包含4个Dof转录因子结合序列(A/TAAAG)及多种激素调节与防御相关的顺式元件。实时荧光定量分析显示,接种可可毛色二孢菌显著诱导VvSUC12和VvDof19基因的表达。通过酵母单杂交实验筛选得到与VvSUC12启动子互作的转录因子VvDof19。酵母双杂交实验证实VvDof19具有自激活活性;进一步通过烟草瞬时表达发现Dof19-GFP的相对GUS活性约为对照GFP的9倍,表明转录因子VvDof19能够激活VvSUC12基因的表达。研究结果为深入研究蔗糖转运蛋白在葡萄免疫反应中的功能奠定基础。     

欧盟、美国和日本加工农产品中农药残留评估现状

本文介绍了欧盟、美国和日本评估加工农产品中农药残留的数据要求和试验准则,以及开展膳食摄入风险评估和推荐农药最大残留限量(MRL)的方法,汇总了用于农药残留评估的加工农产品及其分类,分析了加工农产品中农药MRL制定现状,为进一步提高我国相关领域技术提供借鉴。比较分析结果表明,欧盟和美国在农药登记资料要求中明确提出了加工试验数据要求并且制定了相应的试验准则,美国在试验准则中描述了加工农产品中农药MRL的推荐原则;美国和日本制定了一定数量的加工农产品中农药MRL,欧盟建立了适用于监管农药MRL的加工因子数据库;欧盟、美国和日本均在相关法规中规定了加工农产品中农药残留的合规性判定。     

大豆饲料产品中主要抗营养因子含量的检测与分析

本试验旨在应用间接竞争酶联免疫吸附测定(ELISA)法以及色谱法测定不同加工工艺的大豆饲料产品中主要大豆抗营养因子大豆球蛋白、β-伴大豆球蛋白和胰蛋白酶抑制因子以及大豆中蔗糖、棉子糖和水苏糖的含量。采用ELISA试剂盒对国内257份大豆制品中大豆球蛋白、β-伴大豆球蛋白和胰蛋白酶抑制因子含量进行检测与分析;利用离子色谱建立了大豆产品中蔗糖、棉子糖和水苏糖的灵敏、准确的同步测定法,并对国内92份大豆制品中寡糖含量进行检测与分析。结果表明:发酵豆粕、膨化大豆、大豆浓缩蛋白和大豆分离蛋白中大豆球蛋白、β-伴大豆球蛋白和胰蛋白酶抑制因子含量相对较低;本试验所建立的离子色谱同步测定大豆中蔗糖、棉子糖和水苏糖的方法具有灵敏度和准确性高、重现性好的优点,适用于大豆寡糖的测定;通过对实际样品的检测和分析,发现发酵豆粕和大豆分离蛋白中3种大豆寡糖含量均相对较低。本试验所得到不同加工工艺的大豆产品中主要抗营养因子含量的相关数据将为大豆在畜禽饲料中的高效利用提供很好的技术支撑。     

血清中不同甘露(聚)糖结合凝集素浓度的绵羊感染绵羊肺炎支原体后免疫因子表达的变化

为研究中国美利奴羊不同甘露(聚)糖结合凝集素(MBL)浓度感染绵羊肺炎支原体(MO)的免疫因子水平变化,本研究选择血清中MBL高、低浓度的绵羊各6只(感染组和对照组各3只),感染组人工感染MO,分别在人工感染前和感染后不同时间,采用荧光定量PCR法检测血液中血清因子及补体表达水平。结果显示,不同MBL浓度的绵羊人工感染后MBL m RNA水平呈下降趋势,MBL高浓度促炎因子IL-2和IFN-γ的m RNA表达水平较高,抗炎因子IL-4的m RNA表达水平在感染后1 d升高,此后开始下降,而IL-4的m RNA水平在14 d后有所升高;MBL低浓度羊感染后其TNF-α的m RNA水平显著升高,随炎症的缓解,逐渐降低;补体C1和C3的m RNA在感染后表现出不同的变化,MO感染可以激活补体途径。本研究结果表明,低血清MBL浓度与绵羊支原体肺炎具有一定的相关性,MBL不同浓度组之间其IL-2、IL-4、TNF-α、IFN-γ、补体C1、C3水平存在差异,低浓度MBL的绵羊更易发生比较严重的炎症反应。     

Effect of multi-enzymes in combination with a direct-fed microbial on performance and welfare parameters in broilers under commercial production settings

The objective of this study was to determine the response of broilers to the combination of multi-enzymes and direct-fed microbial (DFM) under commercial production settings. A total of 7,000 1-day-old male broilers (Ross 308) were distributed over 10 pens (700 broilers/pen). Two dietary treatments were tested using complete randomized design, including a control diet and a test diet with addition of multi-enzymes (xylanase, amylase, and protease (XAP)] and DFM (a combination of spores from 3 strains ofBacillus amyloliquefaciens). Pelleted diets were offered ad libitum in 3 phases and water was freely available. During starter and grower phases (0 to 21 d), the enzyme and DFM combination resulted in improved FE (P < 0.05). During the finisher phase, higher feed intake and BW gain (P < 0.05) were observed for the test group. Overall, there were significantly higher feed intake, BW gain, and lower water-to-feed ratio in test group compared to the control group. This was related to improved (P < 0.05) modified production efficiency factor which was calculated based on final BW, survival rate, feeding period, and mortality-weight-corrected FCR. The test group had improved litter quality and a reduced foot-pad lesion score compared to the control. In addition, there was a tendency (P < 0.1) of reducingClostridium perfringens population in cecal digesta and higher lactic acid content in the ileal digesta, when expressed on an as-is basis, in the test group. In this study, we demonstrated that using a multi-enzymes and DFM combination in the diet for broilers can result in improved FE in starter/grower phases and animal welfare parameters, and lead to improved production efficiency under commercial settings.     

Genetic characterization of bovine viral diarrhea virus strains in Beijing,China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle

To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5''-untranslated region (5''-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV.