Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

禽胰多肽对肉鸡肝脏和小肠组织中APPR mRNA表达量影响的研究

本试验旨在研究禽胰多肽(APP)对肉鸡肝脏和小肠组织中APPR mRNA表达量的影响.试验以APP粗提液及层析APP制品为材料,选取90只1周龄艾维菌肉鸡随机分为5组,每组3个重复,每个重复6只鸡.Ⅰ设为对照组,Ⅱ、Ⅲ设为饲饮组,Ⅳ、Ⅴ设为注射组.在第7周末,屠宰取其肝脏、小肠组织,运用SYBR Geen Ⅰ实时荧光定量PCR(RT-PCR)法对肝脏和小肠中APPR mRNA的表达进行相对定量分析.结果发现:注射组与饲饮组肝脏及小肠中APPR mRNA的表达水平与对照组相比均呈下降趋势,且注射组降低程度大于饲饮组降低程度;其中,注射组与饲饮组中均是层析APP制品比APP粗提液的下降较明显.提示在外加APP情况下可对APPR mRNA的表达呈现抑制作用.本研究结果为进一步探讨APP与其受体之间的关系,从而为阐明其生理功能和作用机制提供一定的理论依据.     

肝片形吸虫雷蚴与尾蚴的扫描电镜观察

肝片形吸虫的母雷蚴、早期子雷蚴、成熟子雷蚴及尾蚴表被结构经扫描电镜观察：在母雷蚴及早期子雷蚴的口孔周围有乳突分布；雷蚴的围领呈领圈样环绕，围领基部的一侧有一个产孔，产孔的开口处呈唇样的构造，从母雷蚴、早期子雷蚴至成熟子雷蚴的发育，其体表绒毛样构造日趋发达。尾蚴背部密布尖刀形棘，腹部具有许多不规则的突起，其间分布有许多微孔；口吸盘及周围无乳突分布，但在腹吸盘周围分布有排列整齐的指状突起。尾蚴体部与尾部相连处有一环形颈状结构，尾蚴尾干背面为一槽状的皱襞。     

山药酸奶工艺的研制

以新鲜牛奶为原料,加入熟化后的山药汁经处理后通过正交试验,再添加山药果粒加工而研制出一种气味浓郁、口感爽滑、酸甜可口,集酸奶和山药的营养保健于一体的新型发酵饮料.     

Mapping B-cell linear epitopes of NS3 protein of bovine viral diarrhea virus

Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus within the family Flaviviridae. NS3 is one of the immunodominance regions of the BVDV viral proteins. To identify the potential B-cell linear antigenic epitopes within BVDV NS3 region, serial overlapping truncations covering the whole region were expressed and purified, and screened by multistep of Western-blot. We found (₁VCKKITEHERCHVNI₁₅), (₂₀AFFGVMPRGTTPRAPVR₃₆), (₄₆RRGLETGWAYTHQGGI₆₁), (₂₈₁EGDMATGITYASYGYFC₂₉₇), (₄₂₆YSGEDPANLRVVTSQSPYVVVATNAIESGV₄₅₅) and (₄₈₁FIVTGLKRMAVTVGEQA₄₉₇) can be recognized by the BVDV infected bovine serum. These proteins have been confirmed by indirect enzyme-linked immunosorbent assay (I-ELISA). The results of this study might open new perspectives on the structure and antibody–antigen reaction of the non-structural proteins and may aid in the clinical application as well.     

Selection and Evaluation of Indexes Commonly Used to Determine Contamination with T-2 Toxin in Pacific White Shrimp Litopenaeus vannamei by the Grey Relational Method

Abstract

The objectives of the present study were to evaluate the effects of different concentrations of the mycotoxin T-2 toxin in feed on muscle performance in the Pacific white shrimp Litopenaeus vannamei, evaluate indexes of physiological variables that indicate T-2 toxin contamination in the shrimp using the grey relational method, and determine the dose–response relationships between T-2 toxin and the indexes. Of the 6 physical, 7 biochemical, and 17 nutritional indexes examined, the values of the grey relational coefficients were highest for the hepatopancreas: body weight ratio (HBR), alanine aminotransferase (ALT) activity, and serine (SER) content (0.83, 0.68, and 0.82, respectively). Therefore, the HBR, ALT activity, and SER content were selected as appropriate indexes for contamination of Pacific white shrimp muscle with T-2 toxin. Based on their dose–response relationship curves, mean effective doses of 1.45, 1.69, and 1.33 mg of T-2 toxin/kg of feed were obtained for the HBR, ALT activity, and SER content, respectively. These results offer technical reference points for the evaluation and control of T-2 toxin in shrimp feed.

Received April 28, 2016 accepted April 9, 2017 Published online July 26, 2017     

绵羊宿营法清除天然植被对土壤和植物的影响

研究绵羊宿营法通过践踏和粪尿对土壤和对天然植被和人工草地的影响。结果表明,绵羊啃食和践踏对植被地上部分损坏严重,8羊夜后草本植物被全部清除,但对土壤容重、紧实度和通气性无显著影响。新鲜粪尿入土后,氨态氮和亚硝态氮浓度迅速上升(最高值分别达377mg·kg^-1和56.1mg·kg^-1),对植物产生毒害作用。单纯尿处理土壤氨态氮和亚硝态氮浓度分别达139mg·kg^-1和28.1mg·kg^-1,显著高于单一粪处理和对照。无毛丑柳叶片粗蛋白质含量显著高于对照(P<0.01)。调查结果表明,羊奶果和黄花香发芽枝条数在试验前后变化不大,但宿营后的无毛丑柳枝条数显著少于处理前。宿营处理的产草量明显高于践踏处理。     

鸡大肠杆菌活的非可培养状态发生相关基因克隆与分析

本研究旨在从分子水平探究细菌活的非可培养状态(VBNC)的发生机制。应用冰乙酸和4℃联合诱导条件,使鸡大肠杆菌进入VBNC状态,并利用mRNA差异显示技术(DDRT-PCR)获得VBNC相关基因。结果表明,从VBNC大肠杆菌中筛选得到的3个差异片段与大肠杆菌23S核糖体RNA基因序列具有较高的核苷酸同源性,分别为98%、98%和99%,而氨基酸的同源性也均在97%以上,表明这3个序列是大肠杆菌23S核糖体rRNA基因的部分序列,同时也是与大肠杆菌VBNC状态发生密切相关的基因。由此推知,当正常大肠杆菌在未暴露任何压力下时,其转录水平较低,特别是23S rRNA的某一(些)基因不显示或受到强烈抑制。当进入VBNC状态后,面临生存压力时,这一(些)基因转录水平明显强于正常状态,而核糖体作为蛋白质合成的主要结构与场所,其某些基因也将积极参与新蛋白质的生物合成。     

2例犬死亡原因的实验室诊断

对昆明某犬场2例死亡犬的死亡原因进行实验室鉴定。采集犬肠内容物分别做细菌和病毒检测。细菌检测包括:犬肠内容物划线培养,纯化,分离,革兰氏染色,镜检,生化试验,血清试验,药敏试验。病毒检测包括:提取病料的总DNA,用犬细小病毒的特异引物进行PCR扩增,扩增产物经胶回收后克隆到pMD18-T载体,转化受体菌DH5α,挑取阳性克隆质粒进行序列测定,并与已知参考毒株序列进行比对,分析核苷酸的同源性。分离到一株有部分耐药性的侵袭性大肠埃希菌;检测到一株犬细小病毒,由引物P1/P2扩增的基因与参考毒株CPV-b同源性为98.58%,由引物P3/P4扩增的基因与参考毒株CPV-b同源性为92.1%。结果表明,2例死亡犬的死亡原因分别为侵袭性大肠埃希菌感染和犬细小病毒感染。     

不同牧草在黄牛瘤胃内的降解特性

选择装有永久性瘤胃瘘管的云南黄牛3只,用尼龙袋法测定六种牧草干物质(DM)、粗蛋白质(CP)的瘤胃降解率。结果表明,苜蓿草和一年生黑麦草的干物质有效降解率显著高于苜蓿草块、光叶紫花苕、大麦秸、非洲狗尾草(P<0.01),但两者之间无显著差异(P>0.05);苜蓿草蛋白质的有效降解率极显著高于苜蓿草块、一年生黑麦草、光叶紫花苕、大麦秸、非洲狗尾草(P<0.01),且大小顺序为:苜蓿草>苜蓿草块>一年生黑麦草>光叶紫花苕>大麦秸>非洲狗尾草。