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Identification of Malassezia Species Isolated from Iranian Seborrhoeic Dermatitis Patients

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Identification of Malassezia Species Isolated from Iranian Seborrhoeic Dermatitis Patients European Review for Medical and Pharmacological Sciences 2010; 14: 63-68 Identification of Malassezia species isolated from Iranian seborrhoeic dermatitis patients M.T. HEDAYATI, Z. HAJHEYDARI*, F. HAJJAR**, A. EHSANI§, T. SHOKOHI#, R. MOHAMMADPOUR‡ Department of Medical Mycology and Parasitology; *Department of Dermatology; **Department of Medical Mycology and Parasitology, School of Medicine, Sari (Iran) §Department of Dermatology, School of Medicine, Tehran University of Medical Sciences, Tehran (Iran) #Department of Medical Mycology and Parasitology, School of Medicine; ‡Department of Statistics, School of Health, Mazandaran University of Medical Sciences, Sari (Iran) Abstract. – Background and Objectives: Key Words: In recent years, the genus Malassezia has come Seborrhoeic dermatitis, Malassezia species, Tween to be considered important in the etiology of se- assimilation. borrhoeic dermatitis (SED). The aim of present study was identification of Malassezia species on the lesions of Iranian SED patients. Methods: 100 patients with SED were en- rolled in the study. The patients were evaluated both clinically for the severity of SED and micro- scopically for the presence of the yeast Malassezia. Diagnosis of Malassezia was made Introduction after the yeast Malassezia was microscopically observed on skin scales stained with methylene Yeasts of the genus Malassezia, formerly blue. All samples were also cultivated on Leem- known as Pityrosporum, are ubiquitous skin resi- ing and Notman and Sabouraud’s dextrose agar culture media. The agar plates were incubated at dents of humans and other warm-blooded animals 32°C for 2 weeks and evaluated for the existence and also associated with several dermatological of growth every day for one week. Identification disorders. In recent years, the genus Malassezia of isolated yeast was based on morphological has come to be considered important in the etiolo- and physiological characteristics. gy of seborrhoeic dermatitis (SED)1-3. The im- Results: From 100 patients with SED, 60% provement of SED (and the related condition, were female. The age range was 12-65 years with dandruff) by treatments directed against median 27.3 years. The highest prevalence of SED was seen in 20-29 years age group. 59% and 41% Malassezia supports the role of the yeasts in of patients had local and generalized lesions, re- these conditions, although other factors such as spectively. 58% of patients showed lesion on an impaired barrier function are also thought to scalp. Microscopic examination of skin scales be important4. So the epidemiology and the ecol- was positive in 100% of SED lesions. 96% of pa- ogy of Malassezia species in SED has been con- tients showed more than 1-3 yeasts in each mi- sidered by different investigators1-3,5-8. Epidemio- croscopic field whereas only 4% patients showed 1-3 yeasts in whole slide. Totally, 77% of the spec- logical data suggest geographical variations in imens yielded Malassezia in culture. Malassezia the rate of the isolated Malassezia species from globosa was the most commonly isolated SED1-3,5-8. Malassezia species (55.8%). Malassezia globosa The taxonomy of Malassezia has undergone had also most frequencies on scalp and face le- extensive revisions in the last 10 years and is still sions. Malassezia furfur had most frequency on in a state of flux. Guillot and Gueho described trunk lesions. and named 7 species of Malassezia (M.): M. fur- Conclusion: The results of our study showed high recovery rate of Malassezia species on le- fur, M. sympodialis, M. obtusa, M. globosa, M. 9 sions of patients with SED. So it might be play- restricta, M. slooffiae and M. pachydermatis . ing a causative role in the etiology of this dis- Subsequently, other species have been described, ease. including M. dermatis10, M. japonica11, M. Corresponding Author: Mohammad T Hedayati, PhD; e-mail: [email protected] 63 M.T. Hedayati, Z. Hajheydari, F. Hajjar, A. Ehsani, T. Shokohi, R. Mohammadpour nana12, M. yamotoensis13, and M. equi, although Species Identification the latter has not been formally recognized as a We identified Malassezia species according to species14. Guillot et al.17. Isolates from the primary cultures In Iran, only a few culture studies have been were used for identification. M. pachydermatis is made using the new classification of the able to grow on Sabouraud’s agar. Tween assimila- Malassezia yeasts15,16 but clinical studies are tion and catalase test were applied for identification lacking. Therefore, it is important to look at the of other Malassezia species. Briefly, Malassezia distribution of the various Malassezia species on yeast suspensions were prepared in sterile distilled the skin of patients with Malassezia associated water, which were adjusted to McFarland No. 2 tur- skin diseases including SED patients. In our bidity, mixed with Mycobiotic agar with cyclohexa- study we present some informations about the mide and chloramphenicol, and poured into the distribution of Malassezia species on the lesions plates. Four holes were made in the agar by means of Iranian SED patients. of a 3-mm diameter punch and filled with 30 μl each of tween 20, 40, 60 and 80, respectively. The agar plates, incubated at 32°C for 1 week, were ex- amined each day for the existence of any growth Materials and Methods around the wells that contained tween compounds. Patients Catalase Test 100 patients with SED from the Department of One drop of hydrogen peroxide solution was Dermatology, of Boali Hospitals (Sari City) and applied onto a yeast colony, which was on a glass from the Department of Dermatology, Hospital slide. Production of gas bubbles was considered Razi, Tehran, were enrolled in the study. The pa- a positive catalase reaction. tients filled out the consent form to participate in research which was approved by the Ethical Statistical Analyses Committee of Mazandaran University of Medical Chi-square test was performed using SPSS Sciences. The severity of the SED lesions was software (version 13.0) and differences were evaluated using a semiquantitative scale of 0 to 3 considered significant at p<0.05. for each of the symptoms (erythema, scaling, itching and seborrhoea), where a value of 0 cor- responded to the absence of symptoms, 1 to mild symptoms, 2 to moderate symptoms and 3 to se- Results vere symptoms. The sum of these values is re- garded as score of SED. 60% of the patients with SED were female. The age range was 12-65 yrs with median and Sampling and Microbiological Evaluation SDF yrs of 27.3 and 10.2, respectively. The high- Sampling was performed from the lesions af- est prevalence of SED was seen in patients with ter diagnosis of SED. Skin scales were scraped 20-29 yrs old. 59% and 41% of patients had local off with a sterile blade and transferred to the lab- and generalized lesions, respectively. 58% of pa- oratory. Diagnosis of Malassezia was made after tients showed lesion on scalp (Table I). budding yeast cells that were microscopically ob- served on skin scales stained with methylene blue. To counting of yeast cells, each slide was Table I. Distribution of SED patients on the basis of age examined under high power field (hpf) of micro- and gender. scope and it was recorded as number of observed yeast cells per hpf. All samples were also culti- Total Male Female Age vated on Leeming and Notman medium and n (%) n (%) n (%) Yrs Sabouraud’s dextrose agar culture media. The 24 (24) 11 (28.9) 13 (21) 10-19 agar plates were incubated at 32 °C for 2 weeks 44 (44) 14 (36.8) 30 (48.4) 20-29 and evaluated for the existence of growth every 16 (16) 6 (15.8) 10 (16.1) 30-39 day for one week. Identification of isolated yeast 13 (13) 7 (18.4) 6 (9.7) 40-49 was based on morphologic and physiologic tests, 3 (3) 0 (0) 3 (4.8) 50≤ namely tween assimilation profiles and catalase 100 (100) 38 (100) 62 (100) Total reaction. 64 Identification of Malassezia species isolated from Iranian seborrhoeic dermatitis patients A: 1-3 yeast cells in whole slide B: 1-3 yeast cells per hpf C: 4-7 yeast cells per hpf D: > 7 yeast cells per hpf Figure 1. Distribution of observed yeast cells number in direct microscopic examination (DME) on lesions of patients with SD. Direct microscopic examination (DME) of Table IV shows the frequency of observed specimens was positive in 100% of SED lesions. yeasts number in DME on lesions of patients Figure 1 shows the observed yeast cells number with SED based on age groups. 48% and 49% of in DME on lesions of patients with SED. 96% patients in 20-29 years age group showed 4-7 (96/100) of patients showed more than 1-3 yeast and >7 yeast cells per hpf, respectively. There cells per hpf whereas only 4% (4/100) patients was significant difference between observed showed 1-3 yeast cells in whole slide. yeasts number in DME on lesions of patients Totally, 77% of the specimens yielded with SED and age groups (p=0.003). Malassezia in culture. The most commonly iso- Out of 100 patients with SED, 9, 74 and 17 pa- lated Malassezia species was M. globosa tients showed mild, moderate and severe SED, re- (55.8%), followed by M. furfur (32.5%), M. re- spectively (Table V). With respect to the observed stricta (9.1%), M. sympodialis (1.3%) and M. yeasts number in DME on lesions, 22.2%, 56.7% japonica (1.3%). M. globosa was most common and 41% of patients with mild, moderate and se- isolated species from face (17/27, 63%) and vere SED had a density of >7 yeast cells per hpf, scalp (25/45, 55.5%). M. furfur had most fre- respectively (Table V).
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