48株临床分离假丝酵母菌DNA异质性分析

应用随机扩增多态性DNA技术(RAPD)对48株假丝酵母菌临床菌株进行PCR扩增,并对扩增产物的指纹图谱进行分析,结果显示:48株假丝酵母菌中白假丝酵母菌35株,非白假丝酵母菌13株,引物1和引物2将来源不同的35株白假丝酵母菌分成4型和6型,且RAPD技术可将白假丝酵母菌鉴定至种,并可分辩同种菌的不同型别,是假丝酵母菌致感染、追踪病原的分子流行病学研究有效方法。     

假丝酵母属疑难菌株大亚基rDNA D1/D2区域序列分析及其分类学意义*

对根据常规形态和生理生化性状难以确定分类学地位的8株假丝酵母菌,进行了以大亚基(26S) rDNA中D1/D2区域(约500~600 bp)的碱基序列分析为依据的分子分类学研究。根据系统树上所显示的供试菌株与假丝酵母属及相关子囊菌酵母已知种的亲缘关系,以及与最近缘种模式菌株D1/D2区域序列的相似性比较,确定了各个菌株的归属。本研究也显示了DNA序列分析在假丝酵母菌快速鉴定中的优越性。     

假丝酵母属疑难菌株大亚基rDNA D1/D2区域序列分析及其分类学意义

对根据常规形态和生理生化性状难以确定分类学地位的8株假丝酵母菌,进行了以大亚基(26S) rDNA中D1/D2区域(约500～600 bp)的碱基序列分析为依据的分子分类学研究.根据系统树上所显示的供试菌株与假丝酵母属及相关子囊菌酵母已知种的亲缘关系,以及与最近缘种模式菌株D1/D2区域序列的相似性比较,确定了各个菌株的归属.本研究也显示了DNA序列分析在假丝酵母菌快速鉴定中的优越性.     

不同培养基对白假丝酵母菌生物膜芽管形成的影响

目的评估不同培养基对产生物膜白假丝酵母菌芽管产生的影响。方法收集临床产生物膜的白假丝酵母菌20株,购买3株可以产生物膜的质控菌株。比较23株白假丝酵母菌在5种不同培养基(人血清、胰蛋白胨大豆肉汤、脑心浸液肉汤、RPMI 1640、NaHCO₃溶液)的出芽菌株数量和出芽率。结果在23株产生物膜的白假丝酵母菌中,人血清和脑心浸液肉汤培养基中23株(100.0％)全部产生芽管,NaHCO₃培养基18株(78.3％)、胰蛋白胨大豆肉汤培养基16株(69.6％)和RPMI 1640培养基15株(65.2％)。芽管生成试验阳性的白假丝酵母菌中,在5% CO₂培养条件下的酵母细胞出芽率更高(孵育2 h,出芽率78.0%),芽管长度更长。结论 脑心浸液肉汤可以代替人血清作为芽管试验的诱导剂,人血清(5% CO₂)可以促进芽管的生成以及延伸芽管的长度。     

复发性外阴阴道假丝酵母菌病菌种分型及药敏

目的对RVVC、VVC患者阴道分泌物进行培养、菌种分型及药物敏感试验,探讨RVVC发生的原因。方法用沙保弱琼脂培养基培养阴道分泌物,分离纯化菌株;VITEK 2全自动微生物分析仪鉴定菌种;同时进行药物敏感试验。结果共分离出101株假丝酵母菌,其中RVVC组45株,VVC组51株,健康组5株。101株中白色假丝酵母菌86株,占85.1%(86/101),RVVC组45株中35株为白色假丝酵母菌,占77.8%(35/45);VVC组51株中47株为白色假丝酵母菌,占92.2%(47/51),健康组5株中4株为白色假丝酵母菌,占80%(4/5)。RV-VC组非白色假丝酵母菌比例高,与VVC组比较差异有显著性(P<0.05)。RVVC组假丝酵母菌对唑类药物敏感性低于VVC组。结论VVC、RVVC的主要致病菌仍是白色假丝酵母菌,RVVC组非白色假丝酵母菌比例高于VVC组。RVVC组假丝酵母菌对氟康唑的敏感率明显低于VVC组。     

深红酵母相关菌株全细胞长链脂肪酸的分析及其数值分类

从黄海和渤海海水中分离到15株红酵母,初步定名为深红酵母(Rhodotorula rubraLodder)对这些菌株及另外8株红酵母属对照种的形态、生理生化性状以及全细胞长链脂肪酸的组成进行了测定,并用多元统计方法对菌株间的相似性进行了计算,对菌株分群。结果表明,15株酵母之间性状存在许多差异,这些差异不能完全被鉴定性状所反映,在多元分析中15株菌不能形成紧密的聚类群。研究结果对利用个别形态和生理生化性状确定的深红酵母种的范围提出了异议。     

假丝酵母属疑难菌株大亚基rDNAD1／D2区域序列分析及其分类学意义

对根据常规形态和生理生化性状难以确定分类学地位的8株假丝酵母菌,进行了以大亚基（26S）rDNA中D1/D2区域（均500-600bp)的碱基序列分析为依据的分子分类学研究。根据系统树上所显示的供试菌株与假丝酵母属及相关子囊菌酵母已知种的亲缘关系,以及与最近缘种模式菌株D1/D2区域序列的相似性比较,确定了各个菌株的归属,本研究也显示了DNA序列分析在假丝酵母菌快速鉴定中的优越性。     

口腔黏膜真菌鉴定方法的比较

比较常见用于黏膜真菌菌种鉴别的多种方法,探寻最佳的鉴别方法。采集230例普通人群口腔黏膜样本,分别用玉米吐温-80培养观察厚膜孢子法、糖发酵生化反应法、CHROMagar假丝酵母菌显色培养基法、ITS基因的PCR-RFLP(聚合酶链反应-限制性片段长度多态性)法、ITS测序菌种鉴定法,鉴别真菌各菌株。结果显示:有56例菌株至少通过1种方法检出真菌;玉米吐温-80分离培养假丝酵母菌37株;50例菌株ITS基因测序共鉴定出8个菌种,白假丝酵母菌(C.albicans)29株,近平滑假丝酵母菌(C.parapsilosis)10株,热带假丝酵母菌(C.tropicalis)5株,Candida metapsilosis 1株,Lodderomyces elongisporus 1株,克柔假丝酵母菌(Candida krusei)1株,乙醇假丝酵母菌(C.ethanolica)1株,季也蒙毕赤酵母菌(Pichia guilliermondii)2株;CHROMagar假丝酵母菌显色培养基法鉴定出3种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌;PCR-RFLP法检出5种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌、季也蒙毕赤酵母菌、克柔假丝酵母菌,与基因的测序鉴定一致率为91%;糖发酵生化反应法阳性标本占被检出真菌例数的46.4%(26/56)。结果表明:ITS基因的测序法可以准确鉴定真菌各个菌种;PCR-RFLP法能鉴定常见的菌种,但操作繁琐;CHROMagar假丝酵母菌显色培养基法能快速准确鉴别3种常见假丝酵母菌菌种;玉米吐温-80可以准确培养鉴别白假丝酵母菌;糖发酵生化反应法,缺乏足够的敏感度和特异性,难以准确鉴别各个菌种。     

Asia1型口蹄疫病毒受体结合位点多样性

摘要：【目的】产甘油假丝酵母作为一株优良高产甘油菌株,已成功应用于工业生产15年。近年来由于产甘油假丝酵母染色体倍性尚不明确,限制了对其进行遗传改造的研究进展,因而我们对产甘油假丝酵母染色体倍性研究,分析确定其染色体倍性。【方法】选用酿酒酵母细胞进行生孢,制备酿酒酵母单倍体细胞作对照,并选用热带假丝酵母作为二倍体酵母细胞对照,利用血球计数板得到热带假丝酵母、产甘油假丝酵母、单倍体及二倍体酿酒酵母细胞数,提取染色体,通过二苯胺检测法测定DNA含量。由于在相同紫外照射条件下单倍体细胞比二倍体细胞更容易死亡,因     

产甘油假丝酵母(Candida glycerinogenes)染色体倍性分析

摘要：【目的】产甘油假丝酵母作为一株优良高产甘油菌株,已成功应用于工业生产15年。近年来由于产甘油假丝酵母染色体倍性尚不明确,限制了对其进行遗传改造的研究进展,因而我们对产甘油假丝酵母染色体倍性研究,分析确定其染色体倍性。【方法】选用酿酒酵母细胞进行生孢,制备酿酒酵母单倍体细胞作对照,并选用热带假丝酵母作为二倍体酵母细胞对照,利用血球计数板得到热带假丝酵母、产甘油假丝酵母、单倍体及二倍体酿酒酵母细胞数,提取染色体,通过二苯胺检测法测定DNA含量。由于在相同紫外照射条件下单倍体细胞比二倍体细胞更容易死亡,因     

Analysis of fatty acid composition of Candida species by gas-liquid chromatography using a polar column

The fatty acid composition of representative Candida species was examined by gas-liquid chromatography (GLC) using a polar column. The major fatty acids were C14:0, C16:0, C18:0 saturated, C16:1 and C18:1 monoenoic series, with or without C18 polyunsaturated acids (C18:2 and C18:3). In Torulopsis glabrata and Saccharomyces cerevisiae the C18:2 and C18:3 acids were not found, but the C10:0 and C12:0 acids were detected in S. cerevisiae. These results indicated that the Candida genus could be distinguished from Torulopsis and Saccharomyces genera by GLC analysis of fatty acids. Quantitative differences in the fatty acid composition between cells grown at high temperature (37 degrees C) and low temperature (25 degrees C) were found generally in Candida species, and the amounts of C18 polyunsaturated acids (C18:2 and C18:3) increased in the cells grown at 25 degrees C. Each Candida species showed a characteristic profile in fatty acid composition. Determination of the cellular fatty acid composition in Candida species is likely to be useful for the grouping or chemotaxonomy of newer isolates of Candida species.     

光对小麦幼叶脂氧合酶活性及膜脂过氧化作用的影响

选用抗旱型小麦品种陕合６号和水分敏感型小麦品种郑引１号的黄化幼苗为材料,研究了光处理对小麦幼叶脂氧合酶活性和膜脂氧化作用的影响。结果表明：光处理后黄叶变绿,叶片中的ＬＯＸ活性降低,丙二醛含量和叶绿素含量增加,膜透性升高,ＩＵＦＡ升高。ＬＯＸ活性与光抑制过程的恢复,光保护过程及膜脂过氧化作用有关。光诱导产生的膜脂过氧化作用是一种“准膜脂过氧化作用”。     

樟科植物中脂肪酸成分的分布

本文报道了樟科11个属49种种子油和‘种果肉油的脂肪酸成分。其中樟族和木姜子族的各属种(滇新樟除外)种子油脂肪酸成分以 C_(10)、C_(12)和 C_(14)饱和或烯酸为主。而厚壳桂属、润楠和楠属种子油脂肪酸成分主要为棕榈酸、油酸和亚油酸。山苍子、天目木姜子、大果山胡椒、山橙、红脉钓樟、鸭公树和锈叶新木姜子种子油中的 C_(10)、C_(12)和 C_(14)烯酸经分离鉴定均是顺式Δ~4烯酸。     

海、淡水驯化对5种微藻脂肪酸组成的影响

对5种微藻进行了脂肪酸分析及海淡水驯化影响其脂肪酸组成的研究,结果表明：谈水小球藻和海水小球藻的特征脂肪酸均为16：0、16：2、18：0、18：2和18：3;淡水斜生栅藻的特征脂肪酸为16：0、18：1和18：3;海水三角褐指藻的特征脂肪酸为14：0、16：0、16：1和EPA。淡水藻海水驯化和海水藻淡水驯化后,特征脂肪酸的种类不发生变化,但各种脂肪酸的含量有明显变化,驯化后,几种特征脂肪酸及总脂肪酸占细胞干重的比例在蛋白核小球藻、小球藻－1和三角褐指藻SS02品系中均有不同程度的提高,而在斜生栅藻、小球藻－2和三角褐指藻ZS08、XSO3品系中均有不同程度的下降。     

INFLUENCE OF TEMPERATURE ON THE FATTY ACID PROFILE OF LISTERIA MONOCYTOGENES

Variation in the fatty acid profile of two Listeria monocytogenes strains grown at varying temperatures was determined. The fatty acid profiles varied greatly at different temperatures. General decreases in relative percentages of branched and medium chain (up to C16:0) fatty acids and variable changes in long chain fatty acids were found with increasing growth temperature. Individual fatty acid percentages between strains were variable. The relative percentages of unknown long chain fatty acids, detected in both strains at various temperatures, were greatest in Scott A (7.07%) and ATCC 19114 (13.15%) at 35C. Results demonstrated that L. monocytogenes had altered fatty acid profile in response to changes in growth temperature.     

Cellular fatty acid composition of Leuconostoc oenos

The cellular fatty acid composition of 70 lactic acid bacteria was examined by capillary gas chromatography. Fifty-four Leuconostoc oenos strains, including three reference, type strains from the other Leuconostoc spp., nine Pediococcus spp. and two Lactobacillus spp. were studied. Eighteen fatty acids were determined, of which 10 were identified by gas chromatography-mass spectrometry. The relative percentages of the 18 fatty acids of the Leuconostoc strains were analyzed numerically and grouped using the unweighted pair-group method. Results show that four clusters could be defined at r = 0.920, with five strains unassigned. The major fatty acids of the Leuc. oenos strains were found to be palmitic acid (C16:0), palmitoleic acid (C16:1–9), oleic acid (C18: 1–9), vaccenic acid (C18: 1–11), dihyrosterculic acid (C19-cyclopropane-9) and lactobacillic acid (C19-cyclopropane-11). It was mainly on the basis of the amounts of oleic acid and the C19-cyclopropane fatty acids that the strains of Leuc. oenos could be distinguished from each other. This is the first report of the occurrence of dihydrosterculic acid in lactic acid bacteria. For the majority of Leuc. oenos strains, the result obtained with the cellular fatty acid analysis confirmed the phenotypic relationships.     

BIOCHEMICAL COMPOSITION AND FATTY ACID CONTENT OF FILAMENTOUS NITROGEN-FIXING CYANOBACTERIA

The biochemical composition and fatty acid content of twelve strains of filamentous, heterocystous, nitrogen-fixing cyanobacteria have been determined. When grown under diazotrophic conditions, protein, carbohydrate, lipid, and nucleic acids comprised 37–52%, 16–38%, 8–13%, and 8–11% of the dry weight, respectively. The presence of a combined nitrogen source resulted in an increase in the protein content of the cells and a decrease in the levels of lipids and carbohydrates, although biomass productivity was not affected significantly. Biochemical composition also changed during culture growth, with the highest levels of proteins and lipids occurring as the culture entered stationary phase, whereas the highest levels of carbohydrate and nucleic acids were found during the exponential phase. Total fatty acid levels in the strains assayed ranged between 3 and 5.7% of the dry weight. With regard to fatty acid composition, all strains showed high levels of polyunsaturated fatty acids (PUFAs) and saturated fatty acids (SAFAs), with values of 24–45% and 31–52% of total fatty acids, respectively, whereas the levels of monounsaturated fatty acids (MUFAs) were in general lower (11– 32%). Palmitic acid (16:0) was the most prevalent SAFA, whereas palmitoleic (16:1n- 7) and oleic acid (18:1n-9) were the most abundant MUFAs in all the strains. Among PUFAs, γ-linolenic acid (GLA, 18:3n-6) was present at high levels (18% of total fatty acids) in Nostoc sp. (Chile) and at lower levels (3.6% of total fatty acids) in Anabaenopsis sp. The presence of GLA has not been previously reported in these genera of cyanobacteria. The rest of the strains exhibited high levels (12–35% of total fatty acids) of α-linolenic acid (ALA, 18:3n-3). Linoleic acid (18:2n-6) was also present at a substantial level in most of the strains. Eicosapentaenoic acid (EPA, 20:5n-3) was also detected in Nostoc sp. (Albufera). Some filamentous nitrogen-fixing cyanobacteria therefore represent potential sources of commercially interesting fatty acids.     

Formula alpha-linolenic (18:3(n - 3)) and linoleic (18:2(n - 6)) acid influence neonatal piglet liver and brain saturated fatty acids, as well as docosahexaenoic acid (22:6(n - 3)).

Saturated fatty acids can be synthesized de novo and play a role in determining properties of structural membranes. The effect of dietary essential fatty acids, linoleic acid (18:2(n - 6)) and alpha-linolenic acid (18:3(n - 3)), on the saturated fatty acid content of membrane phospholipid has not previously been considered in newborn nutrition. The studies report the effect of low (1% fatty acids) or high (4%) formula 18:3(n - 3) with low (16%) or high (30-35%) formula 18:2(n - 6) on the saturated and unsaturated fatty acid composition of liver and brain structural lipid of piglets fed formula from birth for 15 days. A significant inverse relationship between the formula % 18:3(n - 3), but not 18:2(n - 6), and the liver phospholipid palmitic acid (16:0) was found. This may indicate a possible effect of dietary 18:3(n - 3) on de novo synthesis of 16:0 and requires further investigation. Monounsaturated fatty acids in both liver and brain were significantly lower in response to high 18:3(n - 3) and to high 18:2(n - 6) plus low 18:1(n - 9) in the formula. Liver phospholipid and brain total lipid % docosahexaenoic acid (22:6(n - 3)) were significantly higher when formula containing 4% rather than 1% 18:3(n - 3) was fed, suggesting that 1% 18:3(n - 3) may limit tissue (n - 3) fatty acid accretion. These results suggest that future studies of essential fatty acid requirements, specifically 18:3(n - 3), should consider possible influences on the saturated fatty acids which also play a functional role in tissue structural lipids.     

Chemotaxonomy of heterocystous cyanobacteria using FAME profiling as species markers

The fatty acid methyl ester (FAME) analysis of the 12 heterocystous cyanobacterial strains showed different fatty acid profiling based on the presence/absence and the percentage of 13 different types of fatty acids. The major fatty acids viz. palmitic acid (16:0), hexadecadienoic acid (16:2), stearic acid (18:0), oleic acid (18:1), linoleic (18:2), and linolenic acid (18:3) were present among all the strains except Cylindrospermum musicola where oleic acid (18:1) was absent. All the strains showed high levels of polyunsaturated fatty acid (PUFAs; 41-68.35%) followed by saturated fatty acid (SAFAs; 1.82-40.66%) and monounsaturated fatty acid (0.85-24.98%). Highest percentage of PUFAs and essential fatty acid (linolenic acid; 18:3) was reported in Scytonema bohnerii which can be used as fatty acid supplement in medical and biotechnological purpose. The cluster analysis based on FAME profiling suggests the presence of two distinct clusters with Euclidean distance ranging from 0 to 25. S. bohnerii of cluster I was distantly related to the other strains of cluster II. The genotypes of cluster II were further divided into two subclusters, i.e., IIa with C. musicola showing great divergence with the other genotypes of IIb which was further subdivided into two groups. Subsubcluster IIb(1) was represented by a genotype, Anabaena sp. whereas subsubcluster IIb(2) was distinguished by two groups, i.e., one group having significant similarity among their three genotypes showed distant relation with the other group having closely related six genotypes. To test the validity of the fatty acid profiles as a marker, cluster analysis has also been generated on the basis of morphological attributes. Our results suggest that FAME profiling might be used as species markers in the study of polyphasic approach based taxonomy and phylogenetic relationship.     

Cellular fatty acid composition of Haemophilus species, Pasteurella multocida, Actinobacillus Actinomycetemcomitans and Haemophilus vaginalis (Corynebacterium vaginale)

The fatty acid composition of 35 Haemophilus influenzae strains was found to be grossly similar and characterized by relatively large amounts of 14:0, 3-OH-14:0, 16:1 and 16:0. The three C18 fatty acids 18:2, 18:1 and 18:0 were also present, but in much lower concentrations. This general pattern was also found for most of the other species of Haemophilus examined (H. aegyptius, H. aphrophilus, H. canis, H. gallinarum, H. haemolyticus, and H. parainfluenzae). Small but distinct quantitative discrepancies were detected for H. ducreyi and the haemin-independent species H. paraphrohaemolyticus, H. paraphrophilus and H. suis. Actinobacillus actinomycetemcomitans was found to be indistinguishable from H. influenzae. Pasteurella multocida also exhibited a fatty acid pattern closely related to that of Haemophilus, but could be distinguished by its higher concentration levels of the C18 fatty acids. The fatty acid pattern of H. vaginalis was considerably different from those of the other species examined. This species lacked 3-OH-14:0 and 18:2 and contained small amounts of 14:0 and 16:0, whereas 18:1 and 18:0 were the major constituents.