南京地区1386株临床分离酵母的鉴定及药敏分析

本文旨在研究临床分离酵母的种类及药物敏感性。采用CHROMagar Candida显色培养基和API20CAUX对我院2008年1～12月分离自临床深部真菌感染患者的1386株酵母进行鉴定,并采用Rosco纸片扩散法分析其对两性霉素B、氟康唑、伊曲康唑、制霉菌素、伏立康唑、5-氟胞嘧啶、克霉唑的药物敏感性,数据用Whonet5.4软件分析。结果显示,分离出的1386株酵母中,白假丝酵母占58.95%,热带假丝酵母占15.95%,光滑假丝酵母占15.15%,克柔假丝酵母占2.74%,其他酵母占7.21%。葡萄牙假丝酵母、近平滑假丝酵母、白假丝酵母、热带假丝酵母、光滑假丝酵母和克柔假丝酵母对氟康唑的敏感率分别为95.45%、94.29%、82.99%、79.19%、68.57%和28.95%;白假丝酵母对5-氟胞嘧啶、两性霉素B、制霉菌素、伏立康唑、氟康唑、伊曲霉唑和克霉唑的敏感率分别为93.76%、93.15%、84.58%、84.09%、82.99%、80.05%和78.09%。结果提示,我院深部真菌感染仍以白假丝酵母为主,其次是热带假丝酵母、光滑假丝酵母;白假丝酵母对5-氟胞嘧啶、两性霉素B的敏感率最高;除克柔假丝酵母外的所有酵母对氟康唑的敏感率都很高。     

阴道分泌物酵母样真菌分离培养及其药敏的临床价值

目的了解阴道分泌物酵母样真菌的分离培养及其药敏临床意义。方法从阴道分泌物中分离到的196株酵母样真菌,采用API-20Aux和ATBFUN GUS进行鉴定和药敏试验,所得结果进行统计分析。结果196株酵母样真菌中,分离率最多的前3种是白色念珠菌（80．5％）、葡萄牙念珠菌（9．5％）和光滑念珠菌（5．6％）,酵母样真菌对两性霉素B和制霉菌素敏感率最高,达94．5％和92．6％,5-氟胞嘧啶、咪康唑、益康唑和酮康唑只有43．6％～58．1％。结论真菌性阴道炎主要以白色念珠菌感染为主,感染的酵母菌除了对多烯类药物敏感率高外,对其他抗真菌药有不同程度的下降,应引起关注。     

胶红酵母CYJ03的体外体内抗氧化活性研究

旨在深入研究海洋源胶红酵母CYJ03的抗氧化能力,挖掘其开发成抗氧化产品的潜力.通过测定CYJ03的过氧化氢耐受性,及其发酵上清液、完整细胞和色素提取物3种不同组分的还原力、自由基清除能力和Fe2+螯合能力来评价该菌株的体外抗氧化能力;通过在罗非鱼日粮中添加CYJ03,测定罗非鱼的生长性能以及血清和肝脏的总抗氧化能力(...     

30株临床分离红酵母的药敏试验研究及文献回顾

目的 测定并分析30株临床分离红酵母的体外药敏结果,另外回顾分析249株文献报道红酵母的药敏资料.方法 从临床菌种保藏库纯化鉴定红酵母,使用ATB fungus3方法测定红酵母药敏数据,并检索MEDLINE 20年来红酵母药敏相关文献.结果 体外药敏实验,几乎全部的红酵母菌株对伊曲康唑、氟康唑、伏立康唑均耐药,5-氟胞嘧啶、两性霉素B对红酵母敏感性较好.与文献回顾结果一致.结论 常用唑类抗真菌药对红酵母敏感性差,两性霉素B、5氟胞嘧啶对红酵母具有良好的抑菌活性,可供临床经验用药.     

胶红酵母利用番茄渣和豆粕为原料发酵生产类胡萝卜素的研究

基于降低微生物类胡萝卜素生产成本的考虑,采用番茄渣、豆粕的纤维素酶酶解产物培养胶红酵母,以单位体积发酵液中的总类胡萝卜素浓度增量作为优化目标,先后运用逐步单因素法和均匀设计法系统性地考查了胶红酵母的总类胡萝卜素产量和增量与各个相关因素之间的关系。实验获得的总类胡萝卜素最大产量以及扣除了番茄渣中的类胡萝卜素含量而计算得到的增量分别为12.25 mg/L和10.25 mg/L。实验结果证明设计的生产工艺能够以较低的成本生产出富含类胡萝卜素的饲料,因而是经济可行的。     

临床酵母样真菌的感染特点及耐药性研究

目的了解临床酵母样真菌的感染类型、分布以及耐药情况,为临床诊断治疗提供合理的用药依据.方法采用常规方法进行真菌培养,用科玛嘉显色培养基联合法国生物梅里埃API 20C AUX鉴定条进行鉴定,药敏试验采用微量稀释法.结果864株酵母样真菌中,白色念珠菌619株（71.6%）,其次为热带念珠菌116株（13.4%）和克柔念珠菌47株（5.4%）,非白色念珠菌感染的比例逐年上升（21.5%）.其中,呼吸道标本酵母样真菌检出率最高,达79.7%,其次是消化道为8.8%,泌尿道为4.7%.科室分布依次为干部科、呼吸科、急诊内科、血液科等;白色念珠菌对两性霉素B、5-氟胞嘧啶高度敏感,达90%以上,对氟康唑和伊曲康唑的敏感性有所降低.结论酵母样真菌的检出率与患者基础疾病密切相关;对氟康唑等药物的敏感性有下降的趋势,未发现对4种药物同时耐药的菌株,提示在治疗中,药敏监测是非常必要的.     

特比萘芬对24株黑酵母样真菌的体外药敏试验研究

目的 评价特比萘芬对7个属24株刺盾霉目（Chaetothyriales）黑酵母样真菌体外敏感性.方法 应用美国国家临床和实验室标准研究所（CLSI）的M38-A2方案.菌悬液终浓度为（0.4 ～5）＆#215;104 CFU/mL,30℃孵育5～7d,测定最低有效浓度（MEC）和最低抑菌浓度（MIC）.结果 特比萘芬对24株黑酵母样真菌MEC范围：0.125 ～4μg/mL,MEC90∶2μg/mL,MEC50∶0.25 μg/mL,GM∶0.392 9 μg/mL,特比萘芬对5株暗色真菌100％生长抑制,MIC范围：1～4 μg/mL,MIC50∶2μg/mL,MIC90∶4μg/mL.结论 特比萘芬对刺盾霉目（Chaetothyriales）中的黑酵母样真菌有较强的抑制作用,50％抑菌作用明显.     

国内首例皮瘤丝孢酵母真菌血症分离株的表型及rDNA序列鉴定

目的通过对1例“脂膜炎”患者外周血分离株的表型及分子生物学鉴定,报道国内首例皮瘤丝孢酵母菌血症。方法采集患者的血及鼻咽结节组织标本通过真菌培养获得分离株,经沙氏培养基、马铃薯培养基、科玛嘉念珠菌显色培养后形态学观察及API 20C AUX系统鉴定,最后经PCR扩增rDNA基因测序证实。采用CLSI制定的M27-A2微量稀释法对病原菌株进行7种药物的体外药敏试验。结果分离株通过表型和分子生物学鉴定为“皮瘤丝孢酵母”。药敏结果显示菌株对两性霉素B耐药,氟康唑、伏立康唑、伊曲康唑、5-氟胞嘧啶敏感。卡泊芬净的抑菌浓度较低,而米卡芬净的抑菌浓度相对较高。患者因多种并发症死亡。结论皮瘤丝孢酵母引起的真菌血症为我国首例报道。API 20C AUX酵母鉴定系统可作为鉴定皮瘤丝孢酵母快速而简便的方法。通过分子生物学DNA序列检测分析能鉴定皮瘤丝孢酵母。     

胶红酵母JB401降解脱色三苯甲烷类染料

从烟梗中分离筛选得到1株能够对三苯甲烷类染料高效脱色的微生物,经ITS-5.8S rDNA分析鉴定为胶红酵母,命名为Rhodotorula mucilaginosa JB401。全波长扫描实验结果证实染料的脱色由胶红酵母降解结晶紫引起。为了提高R.mucilaginosa JB401脱色结晶紫的能力,通过单因素试验对R.mucilaginosa JB401的培养条件进行了优化,得出菌体生长24 h后以2%接种量接入初始pH为5的脱色培养基并在37℃摇床培养,可以取得最优脱色效果,此时脱色50、100和200 mg/L的结晶紫达到90%去除率分别需要3、6和14 h。此外,胶红酵母对温度和pH良好的适应性使其具有应用于工业废水处理的潜力。     

红酵母COS—5产胡萝卜素条件的研究

研究碳源、氮源和添加剂对红酵母（Rhodotorulasp．）COS－5产胡萝卜素的影响,并通过正交试验优化其产胡萝卜素的培养基组成。结果表明,COS－5产胡萝卜素的适宜培养基：蔗糖50g、蛋白胨5g、酵母膏5g、桔子皮15g、盐酸硫胺素0．002g、定容1L,pH6．0.250mL三角瓶装培养基30mL。在上述条件下28℃振荡培养96h,细胞生物量为27．5mg干重／mL发酵液,胡萝卜素含量达492．7μg／g干重细胞。COS－5胡萝卜素在474．4nm、338．6nm和310．2nm处有     

一种遗传转化方法在海洋红酵母(Rhodotorula mucilaginosa)中的应用

海洋红酵母作为一种产油微生物富含油脂和类胡萝卜素,但由于没有合适的遗传操作方法,无法对细胞反应器进行理性改进,阻碍了它的进一步发展。首先利用农杆菌介导转化成功构建了海洋红酵母的遗传操作平台。参考完成基因注释的圆红冬孢酵母基因组信息,分离其甘油醛-3-磷酸脱氢酶启动子,以基于圆红冬孢酵母密码子优化的潮霉素作为筛选标记,构建载体转化至农杆菌AGL1中,利用农杆菌介导转化成功得到潮霉素抗性表型正确的转化子,通过表型验证,基因型验证以及Western blot在蛋白水平验证,鉴定了hyg基因的有效表达,证明了外源抗性基因成功导入海洋红酵母菌株中,实现了海洋红酵母新的转化方法的建立。     

Biofilm formation by the yeast Rhodotorula mucilaginosa: process,repeatability and cell attachment in a continuous biofilm reactor

Yeast biofilms contribute to quality impairment of industrial processes and also play an important role in clinical infections. Little is known about biofilm formation and their treatment. The aim of this study was to establish a multi-layer yeast biofilm model using a modified 3.7 l bench-top bioreactor operated in continuous mode (D = 0.12 h^?1). The repeatability of biofilm formation was tested by comparing five bioprocesses with Rhodotorula mucilaginosa, a strain isolated from washing machines. The amount of biofilm formed after 6 days post inoculation was 83 μg cm^?2 protein, 197 μg cm^?2 polysaccharide and 6.9 × 10⁶ CFU cm^?2 on smooth polypropylene surfaces. Roughening the surface doubled the amount of biofilm but also increased its spatial variability. Plasma modification of polypropylene significantly reduced the hydrophobicity but did not enhance cell attachment. The biofilm formed on polypropylene coupons could be used for sanitation studies.     

Identification, antifungal susceptibility and scanning electron microscopy of a keratinolytic strain of Rhodotorula mucilaginosa: a primary causative agent of onychomycosis

Onychomycosis is a dermatological problem of high prevalence that mainly affects the hallux toenail. Onychomycosis caused by the yeast Rhodotorula mucilaginosa was identified using colony morphology, light microscopy, urease and carbohydrate metabolism in a 57-year-old immunocompetent patient from Rio de Janeiro, Brazil. High-resolution scanning electron microscopy of nail fragments, processed by a noncoating method, led to the observation with fine detail of the structures of both nail and fungus involved in the infection. Yeasts were mainly found inside grooves in the nail. Budding yeasts presented a spiral pattern of growth and blastoconidia were found in the nail groove region. Keratinase assays and keratin enzymography revealed that this isolate was highly capable of degrading keratin. Antifungal susceptibility tests showed that the fungus was susceptible to low concentrations of amphotericin B and 5-flucytosine and resistant to high concentrations of fluconazole, itraconazole, voriconazole and terbinafine. These findings showed data for the first time concerning the interaction of R. mucilaginosa in toenail infection and suggest that this emerging yeast should also be considered an opportunistic primary causative agent of onychomycosis.     

Microbiological Characteristics and Susceptibility Patterns of Strains of Rhodotorula Isolated from Clinical Samples

The members of the genus Rhodotorula show a marked ubiquity. In man, they have been isolated from faeces, nails, skin, sputum, digestive tract and adenoids, forming part of the normal human flora, although in recent years cases have been reported of both local and systemic infection by this yeast. There are virtually no studies in the literature on the sensitivity of this genus to the antifungal agents in common clinical use. Therefore, it is considered of interest to study the microbiological characteristics and the susceptibility patterns of Rhodotorula isolated from clinical samples. A total of 35 different strains of Rhodotorula were studied. In vitro susceptibility testing to 5-fluorocytosine, amphotericin B, ketoconazole, fluconazole and itraconazole was performed. All the strains were considered sensitive to 5-fluorocytosine, amphotericin B, ketoconazole and itraconazole and resistant to fluconazole. As a conclusion, we can state that all the antifungal agents tested, except fluconazole, are useful medicaments for the treatment of infections by the Rhodotorula genus. This revised version was published online in June 2006 with corrections to the Cover Date.     

Biochemical characterization and evaluation of invertases produced from Saccharomyces cerevisiae CAT-1 and Rhodotorula mucilaginosa for the production of fructooligosaccharides

Abstract

Invertases are used for several purposes; one among these is the production of fructooligosaccharides. The aim of this study was to biochemically characterize invertase from industrial Saccharomyces cerevisiae CAT-1 and Rhodotorula mucilaginosa isolated from Cerrado soil. The optimum pH and temperature were 4.0 and 70?°C for Rhodotorula mucilaginosa invertase and 4.5 and 50?°C for Saccharomyces cerevisiae invertase. The pH and thermal stability from 3.0 to 10.5 and 75?°C for R. mucilaginosa invertase, respectively. The pH and thermal stability for S. cerevisiae CAT-1 invertase from 3.0 to 7.0, and 50?°C, respectively. Both enzymes showed good catalytic activity with 10% of ethanol in reaction mixture. The hydrolysis by invertases occurs predominantly when sucrose concentrations are ≤5%. On the other hand, the increase in the concentration of sucrose to levels above 10% results in the highest transferase activity, reaching about 13.3?g/L of nystose by S. cerevisiae invertase and 12.6?g/L by R. mucilaginosa invertase. The results demonstrate the high structural stability of the enzyme produced by R. mucilaginosa, which is an extremely interesting feature that would enable the application of this enzyme in industrial processes.