In vitro susceptibility of the seven Malassezia species to ketoconazole, voriconazole, itraconazole and terbinafine

Fifty-five strains, either authentic or ex-type, of seven Malassezia species were investigated for in vitro susceptibility to various concentrations (0.03-64.0 microg/mL) of three azole drugs, ketoconazole, voriconazole and itraconazole, as well as the allylamine terbinafine, using the agar dilution method. All strains of the seven Malassezia species were susceptible to the three azole drugs at low concentrations. M. furfur, M. sympodialis, M. slooffiae, M. pachydermatis, M. globosa, M. obtusa and M. restricta were most sensitive to ketoconazole and itraconazole, with minimum inhibitory concentrations (MICs) ranging from < or = 0.03 to 0.125 microg/mL. The recently introduced antifungal, voriconazole, was also very effective, with MIC80 values < or = 0.03 microg/mL for 80% of strains. MICs of terbinafine against the seven Malassezia species ranged from 相似文献   

花斑糠疹和马拉色菌毛囊炎菌种分布特点分析

目的探讨花斑糠疹和马拉色菌毛囊炎的菌种分布特点。方法从临床诊断为花斑糠疹和马拉色菌毛囊炎的患者分离培养菌种,通过形态学和生理生化学方法鉴定菌种,并比较两组患者菌种分布情况。结果共收集花斑糠疹病例161例,培养阳性121株,其中合轴马拉色菌46株,糠秕马拉色菌13株,球形马拉色菌29株,钝性马拉色菌33株;马拉色菌毛囊炎135例,培养阳性114株,其中合轴马拉色菌52株,糠秕马拉色菌43株,球形马拉色菌13株,钝性马拉色菌6株。两种疾病菌种分布差异有显著性(P<0.005)。结论花斑糠疹和马拉色菌毛囊炎菌种分布存在差异,可能为两种疾病不同临床表现的原因之一,但结果尚需慎重解释,需要分子水平的进一步研究。     

南通和南京马拉色菌毛囊炎临床调查及致病菌种分析

目的 探讨南通和南京马拉色菌毛囊炎的易感因素及致病菌种在不同地区、不同部位马拉色菌毛囊炎中的菌种分布情况。方法 对符合病例收集纳入标准的患者进行问卷调查,取毛囊内容物进行真菌涂片、培养;并根据形态学和生理生化特征进行菌种鉴定。结果 241例临床诊断为马拉色菌毛囊炎的患者,真菌涂片204例阳性,涂片阳性率84.65%;收集标本259份,共得阳性株213株,其中马拉色菌209株,念珠菌4株（占1.54%）,真菌培养阳性率82.24%。菌种鉴定：209株马拉色菌活化菌种后,可供鉴定的马拉色菌菌株186株,共检测到6个菌种的马拉色菌,其中糠秕马拉色菌111株（59.68%）、斯洛菲马拉色菌43株（23.12%）、合轴马拉色菌17株（9.14%）、球形马拉色菌9株（4.84%）、厚皮马拉色菌4株（2.15%）、钝形马拉色菌2株（1.08%）。不同个体、不同部位的菌种分布：胸部、后背、腹部和面颈部以糠秕马拉色菌为主,上肢、肩部和头顶以斯洛菲马拉色菌为主,下肢均为球形马拉色菌。同一个体、不同发病部位存在不同的菌种,主要为糠秕马拉色菌合并合轴或斯洛菲马拉色菌。 结论 南通和南京马拉色菌毛囊炎存在6种马拉色菌致病菌种,糠秕和斯洛菲马拉色菌是主要的致病菌种。     

IgE antibodies to Malassezia furfur, M. sympodialis and Pityrosporum orbiculare in patients with atopic dermatitis, seborrheic eczema or pityriasis versicolor, and identification of respective allergens

Malassezia yeasts may be a trigger factor for atopic dermatitis. Following the recent reclassification of the genus, the presence of specific IgE antibodies was examined in the sera of patients with atopic dermatitis (n = 223), pityriasis versicolor (n = 83), seborrheic eczema (n = 50) and hymenoptera allergy (n = 39) and in controls without skin diseases (n = 50). In addition to using the commercially available radioallergosorbent test (RAST) for Pityrosporum orbiculare couplings were also made against the reference strains for M. furfur and M. sympodialis. To characterize the specificity and molecular weight of corresponding epitopes identical material was used for production of an immunoblot. Despite high total levels of IgE, controls and patients with pityriasis versicolor showed no specific IgE antibodies. Six patients (12%) with seborrheic eczema were positive while 78 patients (35%) with atopic dermatitis had specific IgE antibodies in higher RAST classes that differed between the Malassezia species. The molecular weights of the main antigens of M. sympodialis and M. furfur were determined to be 15, 22, 30, 37, 40, 58, 79, 92, 99 and 124 kDa and 15, 25, 27, 43, 58, 92, 99 and 107 kDa, respectively. Evaluated according to the location of their disease, patients with head and neck lesions most frequently showed Malassezia-specific IgE antibodies. However, there were differences between the Malassezia species tested, the previously used strain P. orbiculare being assignable to the species M. sympodialis.     

Effects of topical vehicles on growth of the lipophilic Malassezia species

In the present study, the abilities of major Malassezia species, M. sympodialis, M. globosa and M. furfur, to assimilate topical agents, which have been widely used as a material of ointment for skin diseases, were tested. Obvious growth of M. furfur on GYEP agar plate was noted in the presence of white petrolatum, purified white petrolatum, hydrophilic ointment and heparinoid in hydrophilic ointment, and also M. sympodialis showed similar growth when they were cultured with hydrophilic or heparinoid in hydrophilic ointment. In contrast, M. globosa did not grow on GYEP in the presence of the any topical agents tested. These results suggest that Malassezia species, especially M. furfur and M. sympodialis, assimilate several topical agents and showed the drug-depended cell growth.     

从包皮龟头炎患者皮损处分离鉴定马拉色菌

目的：了解马拉色菌属各菌种在包皮龟头炎皮损处的构成及其在发病中的作用。方法：从患处取材直接镜检查见马拉色菌孢子和(或)菌丝的包皮龟头炎患者作为研究对象。用胶带法取材后分别接种在含菜子油的培养基及无放线菌酮的沙堡培养基分离菌种。依据生理生化和形念学特点及转种到科玛嘉显色培养基和米粉吐温80琼脂培养结果鉴定出马拉色菌和(或)念珠菌。结果：81例患者中有57例(70．37％)培养并鉴定出马拉色菌(共60株)，其中糠秕马拉色菌20株(33．33％），合轴马拉色菌18株(30．00％)，钝形马拉色菌17株(28．33％)，球形马拉色菌5株(8．33％)。有37例同时分离到念珠菌(其中72．97％为白念珠菌）。44例仅检出马拉色菌的患者中有23例接受抗真菌治疗。结论：糠秕马拉色菌、合轴马拉色菌、钝形马拉色菌是包皮龟头炎患者皮损处的主要菌种；马拉色菌可能单独或与念珠菌协同引起包皮龟头炎。     

从夫妻双方的头皮屑中分离和鉴定马拉色菌

目的 探讨夫妻双方的头皮屑中马拉色菌的带菌情况，菌种构成及夫妻间共用梳子与马拉色菌菌种一致性的关系，方法 采用Leeming和Notman培养基，Dixon培养基和含菜子油培养基分别同时培养114对夫妻的头皮屑，观察培养阳性率与头皮屑严重程度的关系。根据生理生化学及形态学特点鉴定菌种，并比较在这3种培养基中菌落初长时间及对菌种有无选择性。结果 从114对夫妻（共228人0中153人的头皮屑中分离到马拉色菌，阳性率为67.11%。培养阳性率与头皮屑的严重程度有正相关关系，3种培养基共分离到马拉色菌，阳性率为67.11%。培养阳性率与头皮屑的严重程度有正相关关系。3种培养基共分离到459株马拉色菌，从中鉴定出合轴马拉色菌（33.33%)，球形马拉色菌（25.05%)，限制马拉色菌（15.47%)，糠秕马拉色菌（13.73%)和钝形马拉色菌（12.42%)共5个种，3种培养基对菌种无选择性，但在含菜子油的培养基中菌落初长时间最短，同一头皮屑在3种培养基上分离到同一菌种有129人（56.58%)，分离到2种菌的有24人（10.53%)，共用梳子的夫妻双方的头皮屑中菌种相同者（35.96%)显著多于菌种不同者（13.16%)；夫妻间共用梳子者头皮屑中菌种相同者（50.67%)显著多于非共用梳子者（7.69%)。结论 头皮屑中的优势菌种主要为合轴和球形马拉色菌，夫妻双方的头皮屑中马拉色菌菌种一致率较高，而夫妻间共用梳子组的菌种一致率更高，提示马拉色菌可在夫妻之间传播，共用梳子可能是重要的传播媒介。     

Antigenic components of Malassezia species for immunoglobulin E antibodies in sera of patients with atopic dermatitis

Antigenic components of Malassezia furfur, M. globosa, M. restricta, M. slooffiae, and M. sympodialis were studied for immunoglobulin E antibodies in sera of patients with atopic dermatitis (AD). Antigenic components were extracted from Malassezia cells by treatment with beta-mercaptoethanol, referred to as 2-ME extract. CBB staining and lectin blots using Con A, LCA, PHA-E4, PNA or RCA120 showed that the 2-ME extracts contained several species-dependent components that differed quantitatively and qualitatively among the Malassezia species at the protein level. In the Western blot with the 2-ME extracts, of 54 sera of the patients with AD (54 patients), the patients' IgE antibodies most frequently recognized components showing molecular weights of 43-46 kDa for M. slooffiae, 12-22 kDa for M. sympodialis, 35-40 kDa for M. restricta, 45-50 kDa for M. globosa, and of 67-72 kDa for M. furfur, respectively. In the correlative study, in which the total band intensities generated for each extract in Western blot were compared among the Malassezia species, the intensity for M. globosa was well correlated with that for M. sympodialis (r=0.756). In the Western blot inhibition test, the 2-ME extract of M. globosa partially inhibited the reaction of the antigenic components of other Malassezia species with the patient's IgE antibodies. These results indicated that Malassezia species contained both species-specific and common antigenic components at the IgE antibody level.     

API-ZYM系统在部分真菌胞外酶种类分析中的应用

目的探讨API-ZYM系统在真菌胞外酶种类分析中的应用价值。方法 4种马拉色菌(糠秕、钝形、合轴、斯洛菲)、白念珠菌和烟曲霉经纯化培养后用生理盐水最终配成浊度为0.5麦氏单位的菌悬液,使用API-ZYM系统对菌悬液进行19种酶定性测定。结果马拉色菌、白念珠菌和烟曲霉间胞外酶谱有较大差异,但均含有碱性磷酸酶、酯酶C4、类脂酯酶C8、白氨酸芳胺酶、酸性磷酸酶、萘酚-AS-BI-磷酸水解酶;4种马拉色菌胞外酶谱相似,均含有3种酯酶。结论 API-ZYM系统对真菌胞外酶谱的测定可用于菌属间鉴别。     

花斑糠疹和马拉色菌毛囊炎致病菌菌种分析

目的使用限制性内切酶方法鉴定临床花斑糠疹和马拉色菌毛囊炎致病菌菌种分布并比较其差异。方法收集临床花斑糠疹和马拉色菌毛囊炎标本,植于Leeming-Notman培养基。培养阳性菌株提取DNA,扩增其26srDNA片段,用CfoⅠ酶和BstF51酶切。结果共鉴定花斑糠疹阳性菌株128份,其中糠秕马拉色菌49份,合轴马拉色菌23份,球形马拉色菌26份,钝性马拉色菌6份,M.japponica 1份,斯洛菲马拉色菌1份,混合感染标本22份。共鉴定马拉色菌毛囊炎阳性标本70份,其中糠秕马拉色菌43株,合轴马拉色菌5株,球形马拉色菌9株,钝性马拉色菌1株,混合感染标本12份。两种疾病菌种分布差异存在统计学意义(P=0.009)。结论花斑糠疹和马拉色菌毛囊炎致病菌菌种存在一定差异。限制性酶切方法为一种准确、快速的马拉色菌菌种鉴定方法。     

从花斑癣患者的皮损区及非皮损区分离和鉴定马拉色菌

目的 研究花斑癣患者皮损区及非皮损区马拉色菌菌种构成;不同解剖部位、皮损颜色及各菌种的分布情况;患者病情和年龄与菌种构成的关系。方法 用无菌胶带粘取113例花斑癣患者皮损区及非皮损区共629个部位的皮屑,分别接种于含菜子油培养基中分离马拉色菌,用生理生化及形态学方法鉴定菌种。结果 皮损区与相对应的非皮损区马拉色菌分离阳性率无差别,非皮损区额部和胸背部分离阳性率高于上、下肢。共分离到565株马拉色菌,鉴定出合轴马拉色菌(44.78%)、糠秕马拉色菌(32.94%)、球形马拉色菌(11.68%)、钝形马拉色菌(5.84%)及限制马拉色菌(4.76%)共5个种,有27处(5.01%)同时分离到两种菌。皮损区与非皮损区菌种构成无明显差别,限制马拉色菌主要从额部分离出。菌种构成与皮损面积无关,但与皮损颜色和患者年龄有关。皮损颜色与病程无关。结论 花斑癣患者皮损区与非皮损区马拉色菌分离阳性率和菌种构成基本一致,与病情无关,而不同解剖部位、皮损类型及年龄患者的菌种构成有一定差异。     

马拉色菌在新生儿皮肤定植的研究

目的探讨马拉色菌在新生儿皮肤的定植及影响因素。方法采用含菜籽油培养基32℃培养。对拟自然分娩的母亲临产时取阴道分泌物接种于试管斜面,用无菌微孔透气胶带粘取新生儿(8处皮肤,从出生当时到出院)、母亲(胸部和手掌,分娩后第2天)及其护士(手掌,新生儿出生后第2天)皮肤标本,接种于平板,根据菌落形态及生理生化方法鉴定菌种。结果①52份母亲阴道分泌物培养阴性。②15名护士共104份手掌部皮肤标本培养阴性。③104例新生儿(自然分娩和剖宫产各52例)出生当时的皮肤标本培养阴性,但出生后第1天有28例(26.92%)培养出马拉色菌,到第8天上升到59例(56.73%),其中糠秕马拉色菌41例(69.49%)。④从104例母亲皮肤上培养出马拉色菌65例(62.50%),其中糠秕马拉色菌49例(75.38%)。⑤母亲和新生儿124株马拉色菌中菌种相同者92例,一致率为74.19%。⑥新生儿前额和面颊马拉色菌培养阳性率最高。结论马拉色菌在新生儿皮肤定植最早发生在出生后的第1天内,主要部位为额面部,主要菌种为糠秕马拉色菌,存在于母亲皮肤的马拉色菌可能是定植菌种的主要来源。     

Distribution of Malassezia species in pityriasis versicolor and seborrhoeic dermatitis in Greece. Typing of the major pityriasis versicolor isolate M. globosa

BACKGROUND: The expansion of the genus Malassezia has generated interest in the epidemiological investigation of the distribution of new species in a range of dermatoses, on which variable results have been reported from different geographical regions. No data are thus far available from South-east Europe (Greece). OBJECTIVES: To study the distribution of Malassezia species in pityriasis versicolor (PV) and seborrhoeic dermatitis (SD) and to investigate whether polymorphisms in the internal transcribed spacer (ITS) 1 region facilitate detection of M. globosa and M. sympodialis subtypes. METHODS: In total, 109 patients with PV and SD and positive Malassezia cultures were included in the study. Age, gender, primary/recurrent episode, disease extent and clinical form of PV were recorded. ITS 1 polymorphisms of M. globosa and M. sympodialis type and clinical strains were investigated by polymerase chain reaction (PCR)-single-strand conformational polymorphism (SSCP) analysis. RESULTS: Malassezia globosa was the prevalent species isolated from PV and SD either alone (77% and 39%, respectively) or in combination (13% and 18%, respectively) with other Malassezia species. The pigmented form of PV was strongly correlated with the female gender. PCR-SSCP differentiated five subgroups of M. globosa with one being associated with extensive clinical disease. All M. sympodialis isolates displayed a homogeneous ITS 1 PCR-SSCP profile. CONCLUSIONS: Malassezia species isolation rates were in agreement with those reported from South-west Europe. PCR-SSCP of the ITS 1 is useful for highlighting prospective clinical implications of M. globosa subtypes.     

Skin colonization by Malassezia species in neonates: a prospective study and relationship with neonatal cephalic pustulosis

OBJECTIVES: To assess skin colonization by Malassezia species in full-term healthy newborns, to investigate factors associated with colonization, and to look at acnelike cephalic pustulosis associated with this carriage. DESIGN: Samples were obtained from neonates and their mothers 0 to 5 days after birth and again 3 weeks later. Clinical patterns of common acnelike pustulosis were reported as mild (<10 papulopustules), moderate (> or =10 papulopustules), or absent. Direct examination and culture of sample. Identification of yeasts was based on microscopic and physiologic criteria. SETTING: A maternity hospital and the pediatric dermatology unit of a university hospital. PARTICIPANTS: Consecutive series of 102 neonates and their mothers. MAIN OUTCOME MEASURES: Incidence of skin colonization and type of Malassezia species found in neonates and correlation with neonatal cephalic pustulosis (neonatal acne). RESULTS: At the first visit, 11 neonates and 36 mothers had cultures positive for Malassezia. Malassezia sympodialis and Malassezia globosa were preferentially cultured. At 3 weeks, 29 (52%) of 56 neonates and 18 (32%) of 56 mothers had cultures positive for only M sympodialis and M globosa. Breastfeeding was not associated with a higher prevalence of Malassezia carriage in neonates. Malassezia colonization was higher when pustulosis was more severe and M sympodialis was found in pustules. CONCLUSIONS: Malassezia colonization begins at birth and increases in the first weeks of life. A high prevalence of M sympodialis in neonates is noted from birth. Its association with neonatal acne is confirmed. Further investigation is needed to study the role of sebum secretion rate and quality in the neonatal period.     

Speziesdifferenzierung von Hefen der Gattung Malassezia mittels Fourier-Transform-Infrarot-Spektroskopie

BACKGROUND: 83 Malassezia strains (65 wild isolates and 18 reference strains) were differentiated to the species level using conventional methods including morphological and biochemical features. These strains were further analyzed by Fourier transform infrared spectroscopy (FT-IRS).RESULTS: FT-IRS analysis allowed a clear separation of Malassezia strains according to species-specific cluster formation. The main differences were found between Malassezia furfur and other Malassezia species. In addition, within the species Malassezia furfur, a separation in two similar groups could be demonstrated. A disadvantage of FT-IRS is the relatively expensive apparatus. A great advantage is the speed and simplicity of the procedure, producing results within minutes.CONCLUSION: In pityriasis versicolor, Malassezia globosa was the dominant species found in 62% of cases. In addition, Malassezia furfur was found in 60% of dandruff cases.     

中药水煎剂对马拉色菌分离株的体外抑菌实验

目的观察苦参、百部、土槿皮、大风子、白藓皮、蛇床子、黄柏、地肤子及其混合液体外抑制马拉色菌的效果。方法参照M27-A方案中的酵母菌微量稀释法进行药敏试验。结果六种马拉色菌标准菌株对本水煎剂的敏感性:糠秕马拉色菌的MIC最低(0.062 5g/mL),其次为厚皮马拉色菌(0.125g/mL),球形马拉色菌(0.5g/mL),钝形马拉色菌(0.5g/mL),斯洛菲马拉色菌(0.5g/mL),合轴马拉色菌(>0.5g/mL)。结论复方中各单味药间对多株马拉色菌有相互协同作用,对合轴马拉色菌有拮抗作用。     

Malassezia furfur in Infantile Seborrheic Dermatitis

Abstract: Malassezia furfur is important in the pathogenesis of a number of dermatologic diseases including seborrheic dermatitis in adults. It has also recently been suggested that M. furfur might be the etiologic agent in infantile seborrheic dermatitis (ISD). We studied the presence of M. furfur in 21 children with the clinical diagnosis of infantile seborrheic dermatitis. Laboratory analyses showed aberrant patterns of essential fatly acids (EFA) in serum characterized by elevated levels of 18:1w9 and 20:2w6. Samples for M. furfur were taken from the foreheads and chests of children with infantile seborrheic dermatitis at the time of diagnosis, directly after treatment to complete healing, and after 1 year with no signs of infantile seborrheic dermatitis. All the patients were treated topically with borage oil containing 25% gammalinolenic acid (GLA). No reduced growth of M. furfur was seen on contact plates prepared with borage oil. The growth of M. furfur seems not to be related to the clinical symptoms in ISD.     

The prevalence of Malassezia yeasts in patients with atopic dermatitis, seborrhoeic dermatitis and healthy controls

Cultures for Malassezia yeasts were taken from both normal-looking skin and lesional skin in 124 patients with atopic dermatitis, 16 patients with seborrhoeic dermatitis and from normal skin of 31 healthy controls. Positive Malassezia growth was found in fewer patients with atopic dermatitis (56%) than in patients with seborrhoeic dermatitis (88%) or in healthy controls (84%, p<0.01). In the patients with atopic dermatitis, fewer positive cultures were found in lesional (28%) than in non-lesional skin (44%, p<0.05), while positive cultures were found in 75% of both lesional and non-lesional skin of patients with seborrhoeic dermatitis (not significant). M. sympodialis dominated in patients with atopic dermatitis (46%) and in healthy controls (69%). In patients with seborrhoeic dermatitis both M. sympodialis and M. obtusa were cultured in 43%. A Malassezia species extract mixture would increase the possibility of detecting IgE sensitization to Malassezia in patients with atopic dermatitis.     

In vitro activity of phytosphingosines against Malassezia furfur and Candida albicans

Long-chain sphingoid bases, e.g. phytosphingosine, sphingosine and sphinganine, main constituents of the stratum corneum, can strongly inhibit the growth of microorganisms that are known to have undesirable effects on the skin. The aim of this study was to investigate the in vitro activity of different phytosphingosine preparations against Malassezia furfur, and, in comparison, against the common facultative pathogenic yeast Candida albicans. An agar dilution test for minimum inhibitory concentration (MIC) investigation of phytosphingosine base, phytosphingosine lactic acid salt, phytosphingosine HCl, and phytosphingosine glycolic acid salt was carried out using D.S.T. agar containing 2% olive oil and 0.2% Tween 80, to allow growth of the lipophilic yeast. M. furfur growth inhibition in vitro could be achieved only at extremely high phytosphingosine concentrations. Phytospingosine base had the lowest MIC value (mean 6,250 microg/ml, corresponding to 0.63% of phytospingosine in the agar). For the different phytosphingosine salts--lactic acid salt, HCl and glycolic acid salt--4-8 fold higher MIC values were noted. Unexpectedly, there was a growth stimulating effect of Malassezia at lower phytosphingosine concentrations. In comparison, growth of Candida albicans strains was inhibited at phytosphingosine concentrations between 152 and 269 microg/ml.     

Molecular analysis of Malassezia species isolated from hospitalized neonates

Abstract:  Malassezia species are a part of the skin microflora of neonates. Under certain circumstances, they can cause diseases ranging from simple pustulosis to lifethreatening fungemia in newborn infants. Little information is available about the epidemiology of Malassezia species in neonates. In the present study, we successfully isolated Malassezia yeasts from 68.7% of hospitalized neonates. Using the polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFPL), M. furfur (88.06%) was identified as the most isolated species, followed in frequency by M. globosa (10.48%), M. obtusa (0.73%), and M. slooffiae (0.73%). Among the variables studied, only a longer stay in the ward resulted in a higher colonization rate. Using multiple logistic regression, only the type of hospital and ward had some effects on the colonization rate. Our results supported the hypothesis that neonates acquire Malassezia flora through direct contact with their mothers or hospital personnel.