巢湖藻–草–泥堆制有机肥对土壤性质的影响

通过田间试验,研究施用由巢湖取出物蓝藻、水草和底泥高温堆肥制作成的有机肥对作物产量、土壤理化性质及微生物学特性的影响。结果表明:施用藻-草-泥有机肥对大豆、花生有明显增产效应。施肥土壤中总氮、碱解氮、总磷、有效磷、有机质、腐殖酸、阳离子交换量和土壤孔隙度均显著高于未施肥土壤,而容重显著降低。与未种植未施肥处理(T1)相比,种植大豆施肥(T4)和种植花生施肥处理(T5)中微生物生物量碳分别增加116.5%、97.8%,微生物生物量氮增加196.7%、170.6%;T4处理的转化酶、脲酶和过氧化氢酶活性分别提高183%、392%和8.9%,T5处理中相应酶活性分别提高116%、268%、1.01%。相关性分析表明,微生物生物量碳、氮和转化酶、脲酶活性都与多种肥力因子呈显著相关,且转化酶及脲酶活性间存在显著相关性。T4处理中过氧化氢酶活性与肥力因子间无显著相关性,而T5处理过氧化氢酶活性与有效磷、有机质、腐殖酸、微生物生物量碳、微生物生物量氮间呈显著相关。巢湖取出物蓝藻、水草和底泥可作为生产有机肥的良好材料以达到资源化利用目的。     

野外试验中热脉冲探针间距校正方法的应用

土壤热特性是土壤的重要参数之一,热脉冲探针是目前土壤热特性最为常用的测量方法。在用热脉冲探针测量土壤热特性时,探针间距对测量结果的准确性影响非常大。而在野外试验中,由于土壤受到植物根系、土壤冻融、土壤动物活动等的影响,会造成探针间距的变化。因此,探针间距的校正在野外实地测量时十分重要。本研究团队在2013年提出热脉冲探针间距的校正方法,即在温度探针中使用两个热敏电阻,并通过理论公式校正探针初始间距后,得到间距变化后的探针真实间距。将此校正方法应用于野外试验中,分别于夏季和冬季监测土壤中三个深度处(3 cm、8 cm和13 cm)探针间距的变化。结果显示探针间距校正后得到的土壤热特性参数与未校正探针间距时的土壤热特性相比,探针间距校正后得到的土壤热特性参数更加准确可靠;但由于冬季表层土壤存在结冰现象,造成热脉冲探针无法准确测量热特性参数。     

以黄连素/羟丙基-β-环糊精为荧光探针检测金刚烷胺

为了建立一种金刚烷胺(AMD)检测的新方法,以盐酸黄连素/羟丙基-P-环糊精(BRH/HP-β-CD)为荧光探针,将HP-β-CD分别与BRH、AMD相互作用,测定AMD、BRH/HP-β-CD及两者结合的荧光强度变化规律,并验证分析该荧光分光光度法的适用性。结果表明,HP-β-CD与BRH生成1∶1的包合物并显著增强黄连素水溶液的荧光强度,当加入AMD后,BRH/HP-β-CD包合物的荧光强度逐渐减弱,据此成功建立一种以BRH/HP-β-CD为荧光探针的金刚烷胺检测新方法。当AMD溶液浓度在0.05~4.5 mg·L^-1范围内时与荧光猝灭值△F之间呈线性关系,相关系数为0.989 3,检测限(S/N=3)为0.03 mg·L^-1。溶液pH值和常见的药物赋形剂均不会对测量结果造成干扰,将本方法用于盐酸金刚烷胺药片的分析,其回收率在92%~101%范围内,相对标准偏差小于1%,说明该方法能够成功用于金刚烷胺的测定。本研究为以超分子包合物为荧光探针用于金刚烷胺类药物的检测提供了理论依据。     

腹泻犬源大肠埃希菌耐药性以及整合子–基因盒检测研究

【目的】了解腹泻犬源大肠埃希菌Escherichia coli的耐药性以及整合子携带情况。【方法】采用K-B纸片扩散法对30株分离自腹泻犬的大肠埃希菌进行19种抗菌药物的敏感性试验;PCR检测菌株中是否携带Ⅰ、Ⅱ、Ⅲ型整合酶基因,对携带有整合酶基因的阳性菌株进一步检测sul1、qac EΔ1基因以及可变区基因盒携带情况。【结果】30株分离株对19种抗菌药物表现出不同程度的耐药性,共产生18种耐药谱,对阿莫西林、氨苄西林、四环素和多西环素的耐药性较高,耐药率分别为90.00%、83.33%、66.67%和63.33%;对其余药物耐药性较低,耐药率低于14.00%;11株分离株含有Ⅰ型整合酶基因且均携带sul1和qacEΔ1基因,未检出Ⅱ型和Ⅲ型整合酶基因;Ⅰ型整合子阳性菌株中,有2株扩增出1 879 bp的耐药基因盒:dfrA12+orfF+aadA2。【结论】本次检测的腹泻犬源大肠埃希菌对抗菌药物呈不同水平的耐药性;基因盒介导的耐药性与菌株的耐药表型存在部分相关性,大肠埃希菌的耐药性与Ⅰ型整合子存在一定关系。     

Climate change impacts and forest adaptation in the Asia–Pacific region: from regional experts' perspectives

Expert opinions have been used in a variety of fields to identify relevant issues and courses of action. This study surveys experts in forestry and climate change from the Asia–Pacific region to gauge their perspectives on the impacts of climate change and on the challenges faced by forest adaptation in the region, and explores recommendations and initiatives for adapting forests to climate change. There was consensus regarding the impacts of climate change on forest ecosystems and on economic sectors such as agriculture and forestry. Respondents also indicated a lack of public awareness and policy and legislation as challenges to addressing climate change. However, the results indicate differences in opinion between regions on the negative impacts of climate change and in satisfaction with actions taken to address climate change,highlighting the need for locally specific policies and research. The study presents specific recommendations to address issues of most concern, based on subregion and professional affiliation throughout the Asia–Pacific region.The results can be used to improve policy and forest management throughout the region. This research will also provide valuable suggestions on how to apply research findings and management recommendations outside of the AP region. The conclusions should be communicated relative to the level of the research and the target audience,ensuring that scientific findings and management recommendations are effectively communicated to ensure successful implementation of forest adaptation strategies.     

Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype

A new assay for the detection of swine influenza virus (SIV) was developed with a novel nucleic acid probe——Molecular beacon in this study. The specific primers and molecular beacon probes were designed according to the conserved region of H3 and N2 genes of SIV H3N2 subtype. A digital RT-PCR assay was developed for detection of SIV H3N2 subtype. The results showed that SIV H3N2 subtype could be identified simultaneously on this microarry with high sensitivity and reproducibility,which could reach to 10⁶ dilute viruse. The conclusion was that the digital RT-PCR method could analyze quantitatively the RNA templates.On the identification of H3N2 SIV,the digital RT-PCR method was much more scientific than Real-time quantitative PCR method.     

H1、H3亚型猪流感病毒双重实时荧光定量RT-PCR检测方法的建立及应用

为建立一种快速、准确地检测H1和H3亚型猪流感病毒（swine influence virus,SIV）的方法,根据H1和H3亚型SIV血凝素（hemagglutinin,HA）基因保守序列,分别设计2对特异性引物和2条TaqMan探针,建立双重实时荧光定量RT-PCR方法。结果显示,该方法敏感性高,可检测到最低拷贝数为10²拷贝/μL;重复性良好,重复孔Ct值的变异系数均在5%以下;特异性好,除H1和H3亚型SIV外,H4、H5、H7、H9亚型SIV、猪瘟病毒、猪繁殖与呼吸综合征病毒、口蹄疫病毒、伪狂犬病病毒检测均为阴性;在田间样品中成功检出1株H1和1株H3亚型SIV,检出率为1.16%。该方法准确、快速、灵敏、特异,可为H1、H3亚型SIV的快速鉴别检测及流行病学调查提供有效的技术手段。     

马铃薯金线虫的分子检测技术

本研究通过利用随机引物OPK-4对马铃薯金线虫和白线虫及甜菜胞囊线虫进行RAPD-PCR,供试马铃薯金线中5个群体能产生630bp的特异性的片段,将特异性片段进行测序后发现5个不同来源的马铃薯金线虫产生的特异性片段序列完全一致。根据测定的DNA序列设计出特异性探针,可有效地用于马铃薯金线虫的分子检测。     

微囊藻和小球藻携带WSSV量的变化及对水体游离WSSV的影响

研究了铜绿微囊藻（Microcystis aeruginosa）和蛋白核小球藻（Chlorellapyrenoidosa）携带白斑综合症病毒（whitespotsyndromevirus,WSSV）数量变化及对水体游离WSSV量的影响。试验设置微藻高、低密度组和不加微藻的对照组,分别加入等量的WSSV粗提液,于第2、第24、第72和第120小时以实时荧光定量PCR检测藻液上清和沉淀藻体的WSSV数量。结果表明,微囊藻和小球藻可携带少量WSSV,且随时间延长而减少;微囊藻携带WSSV量与其细胞数呈显著正相关（P〈0．05）,小球藻携带WSSV量与其细胞数相关性不显著（P〉0．05）;2种微藻均有促进水体WSSV数量消减的效果,且小球藻的消减效果优于微囊藻,对养殖对虾白斑综合症（whitespotsyndrome,WSS）的生态防控更有积极意义。     

Q热贝纳柯克斯体TaqMan荧光定量PCR检测方法的建立

根据Q热贝纳柯克斯体(Coxiella burnetii)插入IS1111序列设计引物和探针,建立快速检测Q热的TaqMan实时荧光定量PCR方法。以梯度稀释含有目的扩增片段的重组质粒作为标准品,进行定量PCR反应。结果显示,该方法能够检测出10个拷贝数的阳性质粒;标准曲线相关系数为0.995,扩增效率为103%;结核分枝杆菌(M.tuberculosis)、衣原体(C.psittaci)、布鲁氏菌(Brucella.spp)及牛血液的核酸样本特异性检测结果均为阴性。本研究建立的TaqMan荧光定量PCR法灵敏度高、特异性好,对Q热的检测与鉴定中具有良好的应用前景。