实验动物皮肤病原真菌DNA快速少量提取法

PCR技术应用于实验动物皮肤病原真菌检测,方法简单、省时。但是,真菌的DNA提取较为困难。本文推荐一种既简单又经济快速的提取皮肤真菌DNA的方法,并能成功用于实验动物皮肤病原真菌质量检测研究。     

临床标本优势菌种分布及耐药分析

目的了解临床标本中微生物分离率及其耐药性,为微生物检验教学改革提供临床依据。方法对近一年来临床标本的细菌分离情况及耐药性进行回顾性统计和分析,使用SPSS 13.0软件,采用计数资料进行统计描述分析。结果从2 989份标本中分离出644株病原菌,其中G-杆菌占67.7%,G+球菌占23.29%,真菌58株占9.01%。药敏结果显示大多细菌呈现泛耐药性。结论临床感染以G-杆菌为主,G-杆菌又以NFGNB常见,而G+球菌中肠球菌占主要地位。且这些细菌对常用抗菌药物耐药严重。     

Characterisation and expression of phospholipases B from the opportunistic fungus Aspergillus fumigatus

The phospholipase B family (PLB) are enzymes sharing phospholipase (PL), lysophospholipase (LPL) and lysophospholipase-transacylase (LPTA) activities. They have been shown to be important virulence factors in several human fungal pathogens including Candida albicans and Cryptococcus neoformans. Aspergillus fumigatus, a human opportunistic fungal pathogen leading to a high rate of mortality in immunosuppressed patients is known to possess an extracellular phospholipase B activity. In this paper, we report the molecular characterisation of three PLB genes from A. fumigatus (afplb) using degenerate primers in PCR amplification and data from the A. fumigatus genome project. They are expressed at 37 degrees C, and two of them (afplb1 and afplb3) are induced by lecithin. They encode proteins of 633, 588 and 630 amino acids, respectively, presenting together a T-Coffee score of 81. They also possess the amino acid triad responsible for enzymatic activity in the mammalian cytosolic PLA2 and other fungal PLBs. AfPLB1 and afPLB3 are secreted with a cleaved signal peptide. The complete cDNA sequences were obtained by RACE-PCR for the two secreted afPLBs and probably account for the extracellular phospholipase activity previously reported in the culture media of A. fumigatus.     

Ocurrence of the antibiotic producing bacterium Burkholderia sp. in colonies of the leaf-cutting ant Atta sexdens rubropilosa

Fungus garden material from recently established Atta sexdens rubropilosa colonies (6-12 months old) was sampled to detect antibiotic producing microorganisms that inhibited the growth of pathogens of insects and of the fungus gardens but did not affect their mutualistic fungus. A bacterium with activity against the entomopathogenic fungus Beauveria bassiana was isolated from 56% of the gardens tested (n=57) and identified from its biochemical profile and from 16S and 23S ribosomal DNA sequences as a member of the genus Burkholderia. The ant-associated Burkholderia isolates secreted a potent, anti-fungal agent that inhibited germination of conidia of the entomopathogenic fungi B. bassiana, Metarhizium anisopliae, of the saprophytic Verticillium lecanii, and also of a specialist fungus garden Escovopsis weberi. Growth of the ant's mutualist fungus was unaffected.     

Isolation of an alpha-methylene-gamma-butyrolactone derivative, a toxin from the plant pathogen Lasiodiplodia theobromae

Lasiodiplodia theobromae is known as a multi-infectious microorganism that causes considerable crop damage, particularly to tropical fruits. When the fruits are infected by L. theobromae, the typical symptom is the appearance of black spots on the surface of the infected fruit. When injected in to the peel of banana, the culture filtrate of L. theobromae induced formation of black spots. The structure of the isolated compound responsible for this effect was determined to be (3S,4R)-3-carboxy-2-methylene-heptan-4-olide on the basis of analysis of MS, IR, and 1H and 13C NMR spectroscopic data, including HMQC, HMBC, and 1H-1H COSY experiments. The active compound was not only isolated from the culture filtrate derived from potato dextrose medium, but also from the extract of infected peels of bananas.     

君子兰软腐病病原体侵染途径与致病规律的研究

本文报导了君子兰(Clivia miniata)软腐病二株病原体的分离及确定;鉴定了病原体的分类地位,一株为链球菌属的一个新种,定名为君子兰链球菌(Streptococcus miniatus Zhou,Ping et al shao sp.nov),另一株为环状芽孢杆菌(Bacillus subtilis);并对其二病原体的侵染途径与致病规律进行了研究。     

白内障患者术后感染性眼内炎的病原菌分布及CD64指数、血清sTREM1、HMGB1表达的临床意义

摘要 目的：分析白内障患者术后感染性眼内炎（IE）的病原菌分布并探讨分化簇64（CD64）指数、血清可溶性髓系细胞触发受体1（sTREM1）、高迁移率族蛋白1（HMGB1）表达的临床意义。方法：选取2018年1月～2022年7月济南市人民医院眼科收治的125例（125眼）白内障术后IE患者为IE组,选取同期150例（150眼）白内障术后无术后IE患者为非IE组。分析术后IE患者病原菌分布,计算CD64指数和检测血清sTREM1、HMGB1表达。采用多因素Logistic回归分析白内障患者术后IE的影响因素,采用受试者工作特征（ROC）曲线分析CD64指数、血清sTREM1、HMGB1表达对白内障患者术后IE的诊断价值。结果：125例术后IE患者房水或玻璃原液中检测出138株病原菌,革兰氏阳性菌、革兰氏阴性菌、真菌分别占比81.88%（113/138）、16.67%（23/138）、1.45%（2/138）。与非IE组比较,IE组糖尿病、玻璃体溢出比例、CD64指数和血清sTREM1、HMGB1水平升高,手术时间更长（P＜0.05）。多因素Logistic回归分析显示,糖尿病、玻璃体溢出、手术时间延长和CD64指数、sTREM1、HMGB1升高为白内障患者术后IE的独立危险因素（P＜0.05）。ROC曲线分析显示,CD64指数与血清sTREM1、HMGB1联合预测白内障患者术后IE的曲线下面积显著高于单独预测。结论：白内障患者术后IE的病原菌以革兰氏阳性菌为主,CD64指数和血清sTREM1、HMGB1表达与白内障患者术后IE独立相关,可能成为白内障患者术后IE的辅助诊断指标,且三指标联合具有较高的预测价值。     

Pathogenicity of Sporotrichum pruinosum and Cladosporium oxysporum,isolated from the bronchial secretions of a patient,for laboratory mice

In this study we have demonstrated the occurrence of Sporotrichum pruinosum and Cladosporium oxysporum in the bronchial secretions of a patient with a presumptive diagnosis of tuberculosis. This observation coupled with the ability of both fungi to cause infection and elicit tissue responses in experimentally infected mice supported a probable etiologic relationship with the patient which could not be confirmed in the absence of histologic evidence. In vitro some antimycotics were tested against S. pruinosum and C. oxysporum by the agar dilution method. Oxiconazole with a minimum inhibitory concentration of 0.1 g/ml^–1 after 72 h and amorolfine at a concentration of 0.001 g/ml^–1 after 72 h were the most active ones against S. pruinosum and C. oxysporum respectively. It is suggested that the isolation of S. pruinosum and C. oxysporum from patients with bronchopulmonary disorders should be viewed with caution. Clinical and laboratory evaluation of such patients should be done critically before arriving at a firm diagnosis.     

A field study using the fungicide benomyl to investigate the effect of mycorrhizal fungi on plant fitness

Summary The effect of vesicular-arbuscular mycorrhiza (VAM) on the fecundity ofVulpia ciliata ssp.ambigua was investigated at two field sites in eastern England by applying the fungicide benomyl to reduce VAM infection. The application of benomyl at the two sites produced very different results. At one site the application of the fungicide reduced the fecundity of plants whereas at the other fecundity was increased. At the first site the reduction in fecundity was linked to a significant reduction in VAM infection on the sprayed plants. The mechanism of the benefit associated with the VAM infection is however unclear: there was no treatment effect on morphology or on phosphorus inflow. At the second site, where fecundity was increased, there was only a negligible amount of VAM infection amongst the unsprayed plants and it is suggested that the increase in fecundity with the application of benomyl may have resulted from a reduction in infection by other, presumably pathogenic, fungi. The value of VAM fungi to the host plant may therefore not be restricted to physiological benefits. They may also provide protection to the plant by competing for space with other species of pathogenic fungi.     

Electrotransformation of the human pathogenic fungus Scedosporium prolificans mediated by repetitive rDNA sequences

The regions encoding the 5.8S rRNA and the flanking internal transcribed spacers (ITSI and ITSII) from two isolates of the human pathogenic fungus Scedosporium prolificans and one isolate of the taxonomically related species Pseudallescheria boydii (S. apiospermum) were sequenced. The sequences of the two S. prolificans isolates were identical. However, there were minor differences between both species. Phylogenetic analysis of known fungal sequences confirmed a close relationship between S. prolificans and P. boydii. An attempt was made to transform S. prolificans by electroporation using a plasmid vector, pMLF2, bearing the Escherichia coli hygromycin B phosphotransferase gene (hph) under the control of Aspergillus nidulans promoter and terminator sequences. To increase transformation efficiency, the sequenced ribosomal cluster of S. prolificans was used to construct a new vector for homologous recombination.