猴痘病毒PCR检测方法的建立

从GenBank上调取猴痘病毒的基因序列,经过分析,找出了猴痘病毒的特异性靶基因序列F3L片段,人工合成猴痘病毒MPV(AF380138)F3L基因片段(48048-48509bp)并插入质粒,作为病毒检测的模拟阳性模板。根据该序列设计并合成PCR引物,对模拟的阳性模板进行扩增,结果能扩增出与目的片段大小一致的条带。建立了PCR检测方法,经条件优化后,对鸡痘、禽痘、羊痘等14种相似病毒核酸进行PCR扩增,发现具有良好的特异性。PCR方法能扩增出0.3pg的阳性模板,显示建立的PCR方法具有高效、快速、特异、灵敏的特点,可用于口岸猴痘病毒的检疫。     

Soil microbial populations and activities as influenced by legume green fallow in a semiarid climate

Tilled fallow-wheat (Triticum aestivum L.) (F-W) is the most commonly used cropping system in the semiarid Canadian prairie. However, because frequent fallowing degrades soil, a partial fallow with annual legumes as green manure [i.e. ‘green fallow’ (GF)] has been proposed as a soil conserving and more bio-resource efficient alternative to bare fallow. We conducted a 6-year study to compare the influence of four GF-W, a F-W and a continuous W (Cont. W) system, on soil microbial communities, microbial biomass (MB) and activities in an Orthic Brown Chernozem, silt loam (Aridic Haploboroll) at Swift Current, Sask., Canada. The four GF legumes used were black lentil (Lens culinaris Medikus), Tangier flatpea (Lathyrus tingitanus L.), chickling vetch (Lathyrus sativus L.) and feedpea (Pisum sativum L.). They were grown to full bloom and then incorporated with a tandem disk. Analysis of soil taken from 0 to 10 cm depth after growing wheat in the sixth year of the experiment (i.e. 15 months after the most recent legume GF had been turned under) showed that most of the soil biochemical and microbiological attributes assessed were significantly improved (compared to F-W) by increasing cropping intensity (Cont. W), and even more by using the GF systems. The average improvement gained from the four GF legumes relative to F-W, was 385% for number of bacteria, 210% for filamentous fungi, 170% for MB-C, 191% for MB-N, 205% for cumulative C mineralization in 30 days at 21 °C, 202% for dehydrogenase, 171% for phosphatase, and 287% for arylsulfatase activity. The biologically active C and N, when expressed as ratios of MB-C or MB-N to total soil C or N, increased from 1.6 and 2.0% in F-W to 1.9 and 2.6% in Cont. W and to an average of 2.4 and 3.5% in GF-W. A sensitivity analysis (ratio of other treatment values to value for F-W) showed that Cont. W, and the GF-W systems even more, increased all major soil biological attributes tested. Among the GF systems, lentil-W consistently increased sensitivity the most, while Tangier flatpea-W usually increased it the least. The dynamic direct and indirect microbiological attributes were more sensitive indicators of changes in soil productivity than total organic C or N. The sensitivity of the attributes decreased in the sequence: Bacteria>Arylsulfatase>Filamentous fungi≈Cumulative C mineralization≈Dehydrogenase>MB-N>Phosphatase≈MB-C. Compared to the earlier published sensitivities of select physical and chemical attributes of soil quality to these same agronomic treatments, the microbiological attributes proved to be far more sensitive and more responsive to the beneficial influence of legume green fallowing in this semiarid loam.     

玉米中转基因成分筛查策略

根据商业化转基因玉米外源基因元件信息,构建玉米转基因检测分子特征矩阵,并根据元件信息建立转基因玉米的筛查检测策略。结果表明:利用CaMV35S启动子、NOS终止子2个元件的组合,可完成对现有转基因玉米转化事件的筛查检测,检测灵敏度可达到1g/kg。该筛查方法可对玉米中的转基因成分进行快速、高灵敏度的筛查。     

极干旱地区玉米节水灌溉制度的研究

本文根据阿拉善盟地区水资源严重紧缺的现状，对玉米进行了为期３ａ的非充分灌溉试验。经方差分析确认试验成果是可靠的。对国外几种有代表性作物──水模型，进行拟合检验，确认当地条件下，玉米适合于Ｊｅｎｓｅｎ模型。由此用随机动态规划方法进行优化灌溉制度的制定，为指导我国西北极干旱地区的农业生产提供理论依据和技术方法。     

The Shetland Islands scrapie monitoring and control programme: analysis of the clinical data collected from 772 scrapie suspects 1985-1997

There were 574 scrapie positive suspects (histopathological scrapie lesions present) and 198 scrapie negative suspects (histopathological scrapie lesions absent). The greatest number of scrapie cases were recorded in sheep of 2, 3 and 4 years of age which represented 17%, 36% and 23% of the scrapie positive suspects, respectively. The sign sensitivities and specificities for the ten recorded signs were, respectively: pruritus (62%, 42%), ataxia (23%, 74%), hyperaesthesia (32%, 74%), wool loss (25%, 73%), fleece discolouration (29%, 85%), bruxism (23%, 69%), nibbling reflex (17%, 58%), head rubbing (47%, 78%), poll rubbing (25%, 83%). These single signs had poor discriminatory values with likelihood ratios close to one (range 0.89-1.21); combinations of the four signs, pruritus, wool loss, ataxia, hyperaesthesia and emaciation were more discriminatory (range 0.30-4.3). This study covered a time period when bovine spongiform encephalopathy (BSE) might have been introduced into the sheep population on the Shetland Islands via contaminated feed. No temporal changes could be detected in the age structure of the affected animals.     

山西省灰霉菌对啶酰菌胺的敏感性测定

啶酰菌胺是一种较新的用于防治灰霉菌的杀菌剂,国内鲜有灰霉菌对该杀菌剂的抗性报道。为了明确山西省灰霉菌对啶酰菌胺敏感性情况,通过菌落直径法测定了山西省8个地区139株灰霉菌对该杀菌剂的敏感性。结果表明,所采菌株中137株为相对敏感菌株,2株表现为低抗,敏感菌株占98%以上,说明山西省灰霉菌对啶酰菌胺具有较高的敏感性,但啶酰菌胺具有中度抗性风险,且灰霉菌极易产生抗药性,故有必要做好抗性预防工作。     

小波神经网络在生态环境敏感性分析中的应用——以呼和浩特市为例

本文应用小波神经模型对呼和浩特市进行生态敏感性分析,探讨了人工智能方法在生态环境敏感性评价中的应用.结果表明:呼和浩特市郊区和土默特左旗的生态区域为轻度敏感区域,托克托县、清水河县、和林格尔县和武川县为中度敏感区域,各旗县区引起生态环境脆弱的原因有所不同,自然因素仍然是主要原因,但人类生产生活活动是直接推动力.因而对不同地区采取不同生态保护措施和产业结构调整具有重要意义.     

等离子体对ICP-MS调谐结果的影响

[目的]为了提高灵敏度和减少氧化物、双电荷等的质谱干扰。[方法]分析了等离子体参数射频功率、载气流量、采样深度、雾化泵泵速对ICP-MS灵敏度、氧化物、双电荷的影响。[结果]以ICP-MS 7700x为例,给出在标准模式下各参数的最佳范围,即射频功率1400～1 500 W,采样深度7.0～8.5 mm,载气流量1.05～1.15 ml/min,雾化泵泵速0.10～0.15 r/s。[结论]该研究得到ICP-MS 7700x标准模式等离子体参数。     

Latent class analysis of the diagnostic characteristics of PCR and conventional bacteriological culture in diagnosing intramammary infections caused by Staphylococcus aureus in dairy cows at dry off

Background

Staphylococcus aureus is one of the most common causes of intramammary infections in dairy cows at dry off. Reliable identification is important for disease management on herd level and for antimicrobial treatment of infected animals. Our objective was to evaluate the test characteristics of PathoProof ™ Mastitis PCR Assay and bacteriological culture (BC) in diagnosing bovine intramammary infections caused by S. aureus at dry off at different PCR cycle threshold (Ct)-value cut-offs.

Methods

Sterile quarter samples and non-sterile composite samples from 140 animals in seven herds were collected in connection with the dairy herd improvement (DHI) milk recording. All quarter samples were analyzed using BC whereas all composite samples were analyzed with PathoProof ™ Mastitis PCR Assay. Latent class analysis was used to estimate test properties for PCR and BC in the absence of a perfect reference test. The population was divided into two geographically divided subpopulations and the Hui-Walter 2-test 2-populations model applied to estimate Se, Sp for the two tests, and prevalence for the two subpopulations.

Results

The Se for PCR increased with increasing Ct-value cut-off, accompanied by a small decrease in Sp. For BC the Se decreased and Sp increased with increasing Ct-value cut-off. Most optimal test estimates for the real-time PCR assay were at a Ct-value cut-off of 37; 0.93 [95% posterior probability interval (PPI) 0.60-0.99] for Se and 0.95 [95% PPI 0.95-0.99] for Sp. At the same Ct-value cut-off, Se and Sp for BC were 0.83 [95% PPI 0.66-0.99] and 0.97 [95% PPI 0.91-0.99] respectively. Depending on the chosen PCR Ct-value cut-off, the prevalence in the subpopulations varied; the prevalence increased with increasing PCR Ct-value cut-offs.

Conclusion

Neither BC nor real-time PCR is a perfect test in detecting IMI in dairy cows at dry off. The changes in sensitivity and prevalence at different Ct-value cut-offs for both PCR and BC may indicate a change in the underlying disease definition. At low PCR Ct-value cut-offs the underlying disease definition may be a truly/heavily infected cow, whereas at higher PCR Ct-value cut-offs the disease definition may be a S. aureus positive cow.