肉牛源细极链格孢菌的分离鉴定

为鉴定肉牛皮肤病病原及其致病性并筛选其敏感药物,本研究取患皮肤病肉牛病变部位皮屑、毛发和痂皮进行病原菌的分离,并对分离菌株进行了形态学鉴定、ITS序列分析、小鼠致病性试验和药敏试验。结果显示分离到1株形态特征与细极链格孢菌极其相似的菌株(PY2-1-2);经PCR扩增、测序和BLAST序列比对,结果显示分离株(PY2-1-2)的ITS基因序列与细极链格孢菌(MG975630.1)的ITS基因序列的相似性为99%,分离株PY2-1-2的ITS基因序列长度为543 bp,该片段包括ITS1、5.8S rDNA和ITS2的全部基因序列以及18S rDNA和28S rDNA的部分基因序列,提交GenBank(MH656780.1)。根据形态学结合ITS基因序列分析结果确定该分离株为细极链格孢菌。小鼠致病性试验结果显示该菌对小鼠有致病性。药敏试验结果显示:该菌对灰黄霉素、氟康唑、伊曲康唑、酮康唑的最小抑菌浓度(MIC)值分别为0.5μg/mL、4μg/mL、1μg/mL、1μg/mL。本研究为今后进一步探究细极链格孢菌引起的皮肤病诊治提供科学有效的参考。     

Establishment of macro-aggregates and organic matter turnover by microbial communities in long-term incubated artificial soils

The study of interactions between minerals, organic matter (OM) and microorganisms is essential for the understanding of soil functions such as OM turnover. Here, we present an interdisciplinary approach using artificial soils to study the establishment of the microbial community and the formation of macro-aggregates as a function of the mineral composition by using artificial soils. The defined composition of a model system enables to directly relate the development of microbial communities and soil structure to the presence of specific constituents. Five different artificial soil compositions were produced with two types of clay minerals (illite, montmorillonite), metal oxides (ferrihydrite, boehmite) and charcoal incubated with sterile manure and a microbial community derived from a natural soil. We used the artificial soils to analyse the response of these model soil systems to additional sterile manure supply (after 562 days). The artificial soils were subjected to a prolonged incubation period of more than two years (842 days) in order to take temporally dynamic processes into account. In our model systems with varying mineralogy, we expected a changing microbial community composition and an effect on macro-aggregation after OM addition, as the input of fresh substrate will re-activate the artificial soils. The abundance and structure of 16S rRNA gene and internal transcribed spacer (ITS) fragments amplified from total community DNA were studied by quantitative real-time PCR (qPCR) and denaturing gradient gel electrophoresis (DGGE), respectively. The formation of macro-aggregates (>2 mm), the total organic carbon (OC) and nitrogen (N) contents, the OC and N contents in particle size fractions and the CO₂ respiration were determined. The second manure input resulted in higher CO₂ respiration rates, 16S rRNA gene and ITS copy numbers, indicating a stronger response of the microbial community in the matured soil-like system. The type of clay minerals was identified as the most important factor determining the composition of the bacterial communities established. The additional OM and longer incubation time led to a re-formation of macro-aggregates which was significantly higher when montmorillonite was present. Thus, the type of clay mineral was decisive for both microbial community composition as well as macro-aggregation, whereas the addition of other components had a minor effect. Even though different bacterial communities were established depending on the artificial soil composition, the amount and quality of the OM did not show significant differences supporting the concept of functional redundancy.     

Molecular characterization of Ascaridia galli from Bangladesh and development of a PCR method for distinguishing A. galli from Heterakis spp.

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.     

Practical Method of High-Speed Access for Medical Digital Image Sequences

The high-speed access for medical digital image sequences was restricted to special device and complicated system in the past. The authors present a new method of high-speed access for medical digital image sequences which can be performed on general computer by means of the combination of Serial ATA(SATA) and Redundant Array of Independent Disk(RAID).Then they introduce the scheme of system configuration and software program. Lastly the testing result is given and the speediness,cheapness and conveniency of this method are elucidated.     

斜板上流式厌氧污泥床处理低浓度有机废水的试验研究

本文报告了用１６．１升斜板上流式厌氧污泥床在常温下处理低浓度有机废水的启动和运行试验研究。试验使用稀释酒精废水为模拟处理基质，启动阶段仅用７７天时间，装置水力负荷达到２ｍ３／（ｍ３．ｄ），ＣＯＤ去除率达到７８．９％，此时进水ＣＯＤ浓度为７６７．８ｍｇ／Ｌ。在第二阶段试验中，仅用一天时间，装置就恢复了时隔半年的正常运行。以后在温度１２℃以上，ＨＲＴ为１２小时，进水ＣＯＤ浓度在６６０．０～９８２．０ｍｇ／Ｌ范围的１２９天稳定运行中，平均ＣＯＤ去除率为７６．１％。结果表明：采用该工艺处理低浓度有机废水在技术上是可行的，具有启动快，运行稳定，工艺控制简便的优点。水浸没的成组浅层斜板在ＵＡＳＢ／ＩＴＳ反应器中有良好的污泥截留能力和稳流效果，容易生成大颗粒污泥。颗粒污泥最大粒径可达５ｍｍ，ＳＶＩ为１８．２ｍｌ／ｇ。颗粒污泥中以甲烷丝菌为优势菌。     

ITS鉴定木材种类：一例司法鉴定研究实例

本研究选取一例非法收购国家重点保护植物油丹的司法鉴定案作为研究实例,在确认鉴定技术可行的前提下,应用ITS序列分析对待检测的原木进行了分子鉴定。本研究采用CTAB法提取原木DNA,对其提纯后进行ITSrDNA的PCR扩增,再对PCR阳性产物进行克隆、测序,将测序结果进行Blast分析和构建系统发育树。结果表明,送检的47根原木中有9根被鉴定为油丹。本研究采用ITS序列系统发育分析方法,成功鉴定了原木种类,为今后类似司法鉴定案件的审理提供了可靠的鉴定意见。     

秤锤树叶片内生真菌的分离鉴定及其对植株生长的影响

本研究从秤锤树叶片中分离内生菌进行分子鉴定,并接种到无菌组培苗中研究其对植株生长的影响。研究结果显示,从秤锤树叶片中共分离获得7株真菌。扩增出的6个菌株经ITS测序,在NCBI网站上进行基因序列比对,鉴定出其中5株菌株均属于球毛壳菌（Chaetomium globosum）,1株属于子囊菌（As-comycete）。进一步实验表明：分离的菌株多数对组培苗的株高生长影响不大,可初步鉴定其为内生菌,其中的属于球毛壳菌的F和H菌株对无菌苗的株高有明显的促进作用。     

再植枸杞根际真菌群落对长期连作的响应研究

受枸杞自身种植特点和道地产区土地资源限制,连作障碍已成为制约宁夏枸杞产业可持续发展的主要因素之一,导致严重的经济损失和生态问题。前期研究表明,连作能够显著影响再植枸杞根际土壤细菌的群落结构和多样性,但就连作对真菌群落结构和功能的影响目前仍不清楚。本文利用实时荧光定量PCR和高通量测序技术,研究连作对再植枸杞根际真菌群落丰度及多样性的影响。实时荧光定量PCR分析表明,与对照样地相比,连作显著促进再植枸杞根际及非根际土壤中细菌和真菌的丰度。但连作对真菌的促进作用明显高于细菌,导致细菌/真菌比例失衡,使再植枸杞根际及非根际土壤微生物环境偏向于真菌型。对测序结果的分析发现,所测定样地中枸杞根际及非根际土壤中的优势真菌门分别为子囊菌门、担子菌门、接合菌门、壶菌门及球囊菌门,其中连作地再植枸杞根际子囊菌门的相对丰度较对照样地显著降低,而接合菌门的相对丰度则显著增加(P0.05)。FUNGuild真菌功能预测也证实连作显著抑制再植枸杞根际土壤丛枝菌根真菌的相对丰度(P0.05)。基于距离的冗余分析(db-RDA)结果表明,土壤pH、电导率、硝态氮和有效磷含量是影响枸杞根际土壤真菌群落变化的主要因子(P0.05),而土壤硝态氮和有效磷含量则是解释非根际土壤真菌群落变化的主要因子(P0.05)。这些结果说明长期施用化肥可能是改变连作地再植枸杞根际土壤真菌群落结构及枸杞-真菌互作关系的主要因素之一。这一研究结果为理解枸杞连作障碍的形成机制提供理论基础。     

2013年粳稻4t实仓储藏一年后优势菌种的分离与鉴定

稻谷极易受到微生物的侵染,粮食微生物的活动不仅会影响到粮食的安全储藏,导致粮食腐坏变质,还会严重影响食用者的身体健康.采用GB 4789.15-2010对稻谷进行传统培养,并从中分离得到两株优势菌株.对分离出的优势菌株进行形态学观察,观察菌株在不同生长时期的形态及菌丝、孢子形态,初步判定两菌株均为曲霉.同时运用聚合酶链式反应对两菌株的ITS及28S片段进行扩增,扩增得517bp的ITS片度及560 bp的28S片段,将基因序列在Genbank中进行相似性比对,最终确定两菌株为溜曲霉(Aspergillus tamarii)和亮白曲霉(Aspergillus candidus).     

一株酵母菌酿造柿子酒的发酵特性及其序列鉴定

通过三因素二次回归旋转组合试验设计，研究了发酵温度、接种量、发酵液初始pH值对菌株PS2酿造柿子酒发酵特性的影响，得出了柿子酒质量与工艺因素间的回归模型。结果表明：3个因素对柿子酒质量均有极显著影响（p〈0.01），最佳工艺参数是发酵温度为22℃，接种量为10．4%，pH值为3．4。利用ITS1-5．8S—ITS2序列分析鉴定该菌株的分类水平，结果表明菌株PS2为Saccharomyces cerevisiae HA6（登录号SC09327）。