正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

Differential regulation on C5 expression in goose versus mammals by glucose/palmitate provides a potential protection for goose fatty liver

Goose fatty liver is a specific type of nonalcoholic fatty liver that is protected from harmful effects associated with severe steatosis. Our previous findings suggest that suppression of the complement C5 may be relevant, but the mechanism is unclear. Therefore, in this study, we first verified the expression pattern of complement genes (including C5) during goose fatty liver formation and then determined the liver fat content and fatty acid composition by high-performance liquid chromatography (HPLC), followed by selecting the differential metabolites to treat HepG2, goose and mouse primary hepatocytes, aiming to explore the mechanism of C5 and inflammation suppression in goose fatty liver. The data confirmed the suppression of complement genes (including C5) in goose fatty livers. Moreover, fat content was significantly higher in fatty liver versus normal ones, with oleic acid and palmitic acid dominantly accounting for the difference. In line with this, high concentration of palmitate led to down regulation of C5 expression in goose primary hepatocytes whereas upregulation in mouse primary hepatocytes and HepG2 cells. In conclusion, regulation on C5 expression by fatty liver related factors including high level of palmitic acid may contribute to the protection of goose liver from severe hepatic steatosis.     

豚草粗提物对杨树水泡型溃疡病的抑菌作用研究

为解决豚草提取物在林业病虫害防治中的推广与应用问题,以豚草叶片为试验材料,采用温浸-超声波联合提取法提取抑菌活性物质,并开展其对杨树水泡型溃疡病病原菌的室内外抑菌试验。室内抑菌试验表明,粗提物浓度为16 mg/mL时,抑菌率达到73.62%;林间防治试验表明,最小抑菌浓度为 40 mg/mL,抑菌率达到95.14%。通过以上抑菌试验确定豚草粗提物对杨树水泡型溃疡病有较好的抑制效果。     

超高效液相色谱—串联质谱法检测鸡血液样品中喹诺酮类药物残留

旨在建立鸡血液样品中多种喹诺酮类药物残留的超高效液相色谱—串联质谱方法。样品经乙腈提取,经Waters Acquity UPLC BEH C₁₈色谱柱分离,以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱,电喷雾正离子（ESI⁺）模式电离,MRM扫描模式检测。经方法学验证,该方法的线性关系、回收率、精密度均符合要求。应用超高效液相色谱—串联质谱法建立了鸡血液样品中多种喹诺酮类药物同时检测的方法,实现了动物血液样品中12种喹诺酮类药物同时定性、定量分析。     

异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。     

放牧牦牛采食量和几种消化生理指标的测定

在莎草科 +禾本科草地上对放牧条件下牦牛采食量和几种消化生理指标进行测定。结果表明 :牦牛的干物质消化率为 69 3 6%± 5 87% ,放牧牦牛日干物质采食量为 12 2 13g/kg·W0 75,大部分食糜在采食后 60小时左右排出 ,但完全排出要在 5天以后     

肉鸡腹水综合征的心电图学研究进展

近年对肉鸡腹水综合征的研究已取得进一步的认识 ,在研究肺动脉高压的同时更注意具有潜在作用的心脏病变的研究。家禽的心脏传导系统比较特殊 ,它含有一个返回分支束 ,使右房室瓣受传导神经的支配。当右房室瓣不受传导神经的支配时 ,瓣膜功能不全而引起右心室容量过大 ,可介导右心室肥大。心电图描记是研究肉鸡腹水综合征发病机理的有效手段之一。文章系统地阐述了肉鸡心脏的形态结构和传导系统的特点以及几种肉鸡腹水综合征试验模型的心电图学特征、肉鸡的心脏病理变化与心电图学特征之间的关系     

社会林业与五指山的自然保护

五指山自然保护区是世界上现存地势最高的原始热带雨林之一，是我省目前面积最大的自然保护区，也是海南三大河流的发源地。但由于保护区横跨琼中、通什、保亭三县、市，紧邻保护区还生存着众多贫困的黎、苗族村民。因此，保护区与周边社区的联系是十分紧密的，社区的发展也直接影响到五指山热带天然林的保护。海南省林业局中德海南热带林保护和恢复项目在社会林业与五指山森林的保护上做出了有益的尝试，就在全省来说，也是首次对社会林业与森林保护上进行的探索。五指山地区社会林业的研究，对保护海南热带雨林也有着积极的意义。特对社会林业与五指山的森林保护进行了阐述、分析和设想。     

微量元素Se缺乏或过量对雏鸡组织Se含量影响的研究

选用１０２６只１周龄罗斯蛋用雏鸡为试验动物，研究不同日粮Ｓｅ水平对雏鸡组织Ｓｅ含量的影响。结果表明雏鸡的Ｓｅ缺乏或过量均可通过各组织Ｓｅ含量的变化反映出来，但反映的敏感性及临界指示值因组织及发育期不同而异。日粮Ｓｅ水平缺乏或过量时，各组织Ｓｅ含量相应降低或增加。在本试验条件下得出罗斯蛋用雏鸡的Ｓｅ需要量范围为０．１４５×１０￣（－６）～０．４９５×１０￣（－６），其中以０．１９５×１０￣（－６）最佳；日粮Ｓｅ含量≤０．０９５×１０￣（－６）或≥０．９９５×１０￣（－６），雏鸡出现Ｓｅ缺乏症或中毒症。