Carriage of Malassezia spp. yeasts in Cornish Rex, Devon Rex and Domestic short-haired cats: a cross-sectional survey

Carriage of Malassezia spp. yeasts in healthy Cornish Rex cats (CRC) was compared with that in Devon Rex (DRC) and Domestic short-haired (DSH) cats. Samples obtained from the left external ear canal, anus and claw fold of digit III of the left fore foot by swabbing, and the axilla and groin using contact plates, were incubated for yeasts on modified Dixon's agar at 32 degrees C for 7 days. Malassezia species were isolated from 90% of the DRC, but from only 39% of the CRC and 50% of the DSH cats. M. pachydermatis accounted for 121 of 141 Malassezia spp. isolates. Five CRC were colonized by M. pachydermatis alone, one CRC yielded only M. nana, and one cat yielded only M. slooffiae, whereas five CRC were colonized by both M. pachydermatis and M. nana and another yielded M. pachydermatis, M. slooffiae and M. nana. M. nana was primarily isolated from the ear canal, whereas M. slooffiae was most often isolated from the claw. Both the frequencies of isolation and the population sizes of M. pachydermatis at all sites sampled in the CRC were comparable to those of 10 healthy DSH cats. Populations of M. pachydermatis in the left axilla and left and right groin in the CRC were significantly lower when compared with counts in a group of 21 healthy DRC, a breed with very similar coat characteristics but prone to seborrheic dermatitis caused by M. pachydermatis.     

Isolation of Malassezia species from healthy cats and cats with otitis

Lipid-dependent Malassezia species have recently been cultured from veterinary specimens. The identification of Malassezia species isolates from animals is important to clarify the epidemiology of these lipophilic yeasts. Malassezia species were cultured from the external ear canals of 63 out of 99 cats with otitis and 12 of 52 (23%) healthy control cats. The rate of isolation in affected animals versus controls was highly significant (P<0.01). Malassezia pachydermatis was isolated as a pure culture in 33 (45.2%) cats, associated with Malassezia globosa and Malassezia furfur in 20 (50%) and 17 (42.5%) animals, respectively. Three different species were isolated simultaneously in three cats (two cats with M pachydermatis, M globosa and M furfur, one subject with M pachydermatis, M furfur and Malassezia sympodialis). M globosa was isolated as the sole species in two animals. The present work confirms the presence of some lipid-dependent species of Malassezia in both healthy and otitic cats.     

Treatment of Malassezia pachydermatis-associated seborrhoeic dermatitis in Devon Rex cats with itraconazole--a pilot study

Treatment of Malassezia pachydermatis-associated seborrhoeic dermatitis with oral itraconazole was investigated in six Devon Rex cats (DRC). The cutaneous populations of Malassezia were determined using contact plates and a swab-wash method before and after 21 days of pulse treatment with itraconazole (5 mg kg-1 once daily, 7 days on, 7 days off, 7 days on). Before treatment, all cats had greasy seborrhoeic dermatitis involving the axillae, groin, claw folds and palmar and plantar interdigital skin, and two had similar lesions on the ventral neck. After treatment, there was a significant (P<0.05) reduction in overall clinical scores and in scores at all individual sites assessed, except for the interdigital skin (P=0.068). Population sizes of M. pachydermatis in the left and right axillae, left and right groin and palmar interdigital skin were significantly (P<0.05) reduced, whereas the reduction in claw fold counts did not reach significance (P=0.068). The dramatic reduction in yeast counts and an associated marked clinical improvement of the seborrhoeic dermatitis provide important pilot data on the potential value of oral itraconazole in the management of seborrhoeic dermatitis associated with M. pachydermatis in DRC.     

Malassezia spp. overgrowth in allergic cats

A series of 18 allergic cats with multifocal Malassezia spp. overgrowth is reported: atopic dermatitis was diagnosed in 16, an adverse food reaction in another and one was euthanized 2 months after diagnosis of Malassezia overgrowth. All the cats were otherwise healthy and those tested (16 out of 18) for feline leukaemia or feline immunodeficiency virus infections were all negative. At dermatological examination, multifocal alopecia, erythema, crusting and greasy adherent brownish scales were variably distributed on all cats. Cytological examination revealed Malassezia spp. overgrowth with/without bacterial infection in facial skin (n = 11), ventral neck (n = 6), abdomen (n = 6), ear canal (n = 4), chin (n = 2), ear pinnae (n = 2), interdigital (n = 1) and claw folds skin (n = 1). Moreover, in two cats Malassezia pachydermatis was isolated in fungal cultures from lesional skin. Azoles therapy alone was prescribed in seven, azoles and antibacterial therapy in eight and azoles with both antibacterial and anti-inflammatory therapy in three of the cats. After 3-4 weeks of treatment, substantial reduction of pruritus and skin lesions was observed in all 11 cats treated with a combined therapy and in five of seven treated solely with azoles. Malassezia spp. overgrowth may represent a secondary cutaneous problem in allergic cats particularly in those presented for dermatological examination displaying greasy adherent brownish scales. The favourable response to treatment with antifungal treatments alone suggests that, as in dogs, Malassezia spp. may be partly responsible for both pruritus and cutaneous lesions in allergic cats.     

Occurrence, distribution and population size of Malassezia pachydermatis on skin and mucosae of atopic dogs

Aim of the present study was to determine the distribution and quantification of Malassezia yeasts on a wide number of cutaneous sites in atopic dogs by means of a semiquantitative swab technique. A possible relationship between the presence of clinical signs and the occurrence and population size of yeasts was attempted. Forty-one privately owned atopic dogs of different age and breed were sampled. Results were expressed as colony forming units per swab. Malassezia colonies obtained from each plate were counted, scored and typed. All dogs yielded Malassezia pachydermatis from at least one skin area. Yeast population mean size by site was 6.98 (S.D.=3.47) as compared to other body areas. The frequence of isolation was higher from interdigital areas (70.7%), ears (63.4%), nail folds (35.7%), mouth (33.3%), groin (30.9%), conjunctiva and axillae (23.8%), perineum and anus (19%), perianal glands (9.5%). Ears, anus, interdigital areas, perianal glands and groin yielded the largest mycotic amount. M. pachydermatis was the sole species of yeast to colonize canine skin in examined animals. No statistical correlation between the presence of cutaneous alterations and Malassezia isolation was detected. Highest scores were not exclusively found on affected areas, but also on lesion-free sites, demonstrating that atopic animals can be heavily colonized also in apparently healthy areas.     

Isolation of Malassezia pachydermatis and M. sympodialis from the external ear canal of cats with and without otitis externa

The objective of this study was to determine the presence of Malassezia spp. in the external ear canal of cats with and without otitis. Forty-five animals were studied, 20 with and 25 without otitis externa (OE). Cerumen or secretion from external ear canal samples was cultured on modified Mycosel agar and sterile olive oil was added to the surface of the medium before specimen seeding. The isolates were analysed for macro- and micromorphology and identified by catalase tests and on the basis of growth on Tween 20, 40, 60 and 80. Malassezia spp. were isolated from 15 out of 20 (75%) animals with otitis and from 7 out of 25 (28%) cats without OE; the difference between the two groups was statistically significant (P < or = 0.05). Malassezia pachydermatis and M. sympodialis were isolated from 60% (12/20) and 40% (8/20) of cats with otitis, respectively, with no significant difference in the frequency of isolation between the two species. In the microflora of the healthy ear canal M. pachydermatis was significantly more common (6/25, 24%) than M sympodialis (1/25, 4%). The present investigation confirms that M. sympodialis can also act as an aetiological agent of feline OE, and if commercial veterinary laboratories do not use media with added lipids for the isolation of Malassezia spp., this might lead to false-negative results.     

Frequency, body distribution, and population size of Malassezia species in healthy dogs and in dogs with localized cutaneous lesions.

Malassezia species are commensal organisms of human and animal skin that occasionally act as opportunistic pathogens. The lipid-dependent species are associated with human skin disorders, whereas the non-lipid-dependent species (Malassezia pachydermatis) is considered as an opportunistic secondary pathogen affecting the canine skin surface and ear canal. This study evaluated the relationship between Malassezia yeasts, their population size, and the occurrence of skin lesions from healthy and skin-diseased dogs. The efficiency of cytological examination and fungal culture for Malassezia detection was also evaluated. From March 2002 to July 2003, 33 healthy dogs and 54 dogs with pruritic localized skin diseases were examined; skin swabs (1218) were collected from 7 anatomical sites for culture and cytological examination. Malassezia prevalence according to anatomical site and the agreement between cytological results and fungal cultures were statistically analyzed. Differences in mean colony forming unit counts between positive healthy and diseased dogs were evaluated using the Bonferroni test for post hoc pair-wise comparisons. In healthy dogs, Malassezia yeasts were most frequently isolated in the perianal and perioral areas. The frequency of isolation and population size of Malassezia species were higher in dogs with localized dermatitis, especially in affected areas, indicating a role for Malassezia in the occurrence of skin lesions. Malassezia pachydermatis was the species most commonly cultured from the skin and external ear canal of healthy and diseased dogs; isolation of lipid-dependent yeasts from healthy dogs was less frequent. Using fungal culture as the gold standard, cytological examination showed good relative specificity (95%) but very low relative sensitivity (30%).     

Retrospective study: the presence of Malassezia in feline skin biopsies. A clinicopathological study

Malassezia spp. dermatitis, a rare disorder in cats, has previously been associated with immune suppression and internal malignancies. This study evaluates the presence and importance of Malassezia spp. in feline biopsy specimens submitted for histopathological examination. Five hundred and fifty haematoxylin and eosin-stained skin biopsy specimens received for histopathological examination between January 1999 and November 2000 were reviewed. Fifteen (2.7%) submissions contained Malassezia organisms in the stratum corneum of the epidermis or follicular infundibulum. Eleven of 15 cats presented with an acute onset of multifocal to generalized skin lesions. All 11 cats were euthanized or died within 2 months of the onset of clinical signs. Seven cats had dermatopathological changes and clinical signs supportive of paraneoplastic alopecia, and three cats had an interface dermatitis suggestive of erythema multiforme or thymoma-associated dermatosis. Histopathological changes were nonspecific in one cat that was euthanized 2 weeks following onset of severe pruritus and alopecia. In three cats, Malassezia spp. were found in localized sites (two chin, one footpads) and appeared inconsequential to their overall health status. One cat had Malassezia spp. in association with cutaneous demodicosis. These findings suggest that Malassezia yeast in dermatopathological specimens from multifocal or generalized lesions should prompt a thorough clinical work-up for internal neoplasia.     

Frequency of yeasts and dermatophytes from healthy and diseased dogs.

The aim of this study was to investigate the presence of dermatophytes and yeasts in healthy and diseased dogs. A total of 633 samples were collected from 26 healthy animals (104 samples), 131 with dermatitis (343 samples), 74 with otitis (148 samples), and 19 with ocular diseases (38 samples). Cultures from healthy animals were positive for Malassezia pachydermatis in 13.5% (7/52) of samples from skin, 42.3% (11/26) from ear, and 3.8% (1/26) from eye. Fungal growth was observed in 20.4% (70/343) samples from animals with dermatitis. Microsporum canis was the most isolated fungus (n = 39), followed by M. pachydermatis (n = 30) and Malassezia sp. (n = 3). Of the 148 samples from dogs with otitis, 90 (60.8%) were positive for M. pachydermatis, and of the clinical specimens from the conjunctiva of animals with ophthalmic disease, 2.6% (1/38) presented positive cultures for M. pachydermatis. Only 14.3% (2/14) of the positive cultures for M. pachydermatis and 40.9% (9/22) of those for M. canis were positive in the direct exam. Direct exams were positive in 84.3% (70/83) of the culture positive samples from affected ears of dogs with otitis. Malassezia pachydermatis may act as an aggravating factor in the occurrence of cutaneous diseases, or the isolation of M. canis may be associated with the onset of dermatophytosis. Fungal culture, rather than microscopic examination, should be used as the definitive diagnostic test for dermatomycoses and otitis.     

Mycologic disorders of the skin

Cutaneous tissue can become infected when fungal organisms contaminate or colonize the epidermal surface or hair follicles. The skin can be a portal of entry for fungal infection when the epithelial barrier is breached or it can be a site for disseminated, systemic fungal disease. The two most common cutaneous fungal infections in small animals are dermatophytosis and Malassezia dermatitis. Dermatophytosis is a superficial cutaneous infection with one or more of the fungal species in the keratinophilic genera Microsporum, Trichophyton, or Epidermophyton. Malassezia pachydermatis is a nonlipid dependent fungal species that is a normal commensal inhabitant of the skin and external ear canal in dogs and cats. Malassezia pachydermatis is the most common cause of Malassezia dermatitis. The diagnosis and treatment of these cutaneous fungal infections will be discussed.     

An idiopathic facial dermatitis of Persian cats

The clinical and histopathological features of 13 Persian cats which presented with chronic skin disease primarily affecting the face are described. Lesions were characterized by black material adherent to the skin and hair, accompanied by erythema and variable degrees of excoriation. Concurrent ceruminous otitis externa was observed in 7 cases. Histopathological examination of skin biopsy specimens showed marked acanthosis with crusting, hydropic degeneration and dyskeratotic basal epithelial cells, a mixed diffuse superficial inflammatory infiltrate and sebaceous hyperplasia. Malassezia pachydermatis yeasts and various bacteria were isolated from the lesions in some of the cats but in no case was antimicrobial therapy curative. The response to glucocorticoids was variable and often poor. No satisfactory therapeutic regimen could be identified and the cause of the disorder is unknown although a genetic basis is possible.     

Higher incidence of Malassezia pachydermatis in the eyes of dogs with corneal ulcer than in healthy dogs

Malassezia pachydermatis is usually associated with otitis and dermatitis in dogs but it can also cause diseases in other species, including humans. In a human neonatal intensive care unit, M. pachydermatis was isolated from an infant's ocular discharge. Therefore, the aim of this study was to ascertain the presence of Malassezia spp. and its possible consequences in dogs' eyes. This research included 19 dogs with unilateral or bilateral corneal ulcers and 60 healthy dogs. A total of 158 clinical specimens from both the groups were obtained from the conjunctival sac of each eye by a calibrated platinum loop. The samples were placed on Dixon and blood agar, incubated at 35 degrees C, and examined daily for 15 days. Then, the strains were subcultured on Sabouraud agar. Of 22 clinical specimens collected from the eyes with corneal ulcers, five cultures (23%) were positive for M. pachydermatis. Of 16 samples collected from the contralateral healthy eye, cultures were positive in three samples (19%). Three animals had unilateral corneal ulcer and positive cultures for M. pachydermatis in both the eyes. Two dogs had unilateral corneal ulcer and positive cultures for M. pachydermatis in the same eye. However, from the 120 samples of 60 healthy dogs, only four clinical specimens (3%) had positive cultures for M. pachydermatis. The findings of M. pachydermatis, in a considerable percentage of clinical specimens from dogs with corneal ulcer, suggest its possible role at least as an aggravating factor in the pathophysiology of this disease.     

In vitro antifungal susceptibility of Malassezia pachydermatis from dogs with and without skin lesions

Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.     

Risk factors for feline infectious peritonitis in Australian cats

The objective of this study was to determine whether patient signalment (age, breed, sex and neuter status) is associated with naturally-occurring feline infectious peritonitis (FIP) in cats in Australia. A retrospective comparison of the signalment between cats with confirmed FIP and the general cat population was designed. The patient signalment of 382 FIP confirmed cases were compared with the Companion Animal Register of NSW and the general cat population of Sydney. Younger cats were significantly over-represented among FIP cases. Domestic crossbred, Persian and Himalayan cats were significantly under-represented in the FIP cohort, while several breeds were over-represented, including British Shorthair, Devon Rex and Abyssinian. A significantly higher proportion of male cats had FIP compared with female cats. This study provides further evidence that FIP is a disease primarily of young cats and that significant breed and sex predilections exist in Australia. This opens further avenues to investigate the role of genetic factors in FIP.     

Phenotypic characterization and in vitro antifungal sensitivity of Candida spp. and Malassezia pachydermatis strains from dogs

Yeasts of the genera Candida and Malassezia can be found as commensal microorganisms in animals. The main species of importance in veterinary medicine are Malassezia pachydermatis and Candida albicans. The objectives of this study were to conduct a phenotypic characterization and to evaluate the in vitro antifungal sensitivity of strains of C. albicans (n=5), C. tropicalis (n=3) and M. pachydermatis (n=32) isolated from dogs. The phenotyping was based on macro and micromorphological features as well as biochemical analysis. The techniques of microdilution in broth and dilution in agar were used to evaluate the in vitro sensitivity of Candida spp. and M. pachydermatis, respectively. The tested drugs were ketoconazole (KTC), itraconazole (ITC), fluconazole (FLC) and amphotericin B (AMB). The morphological analysis of the strains of Candida spp. and M. pachydermatis did not show any noteworthy alterations when compared to standard strains. On the other hand, in the biochemical tests, 34.4% of the strains of M. pachydermatis were negative for the urease test. Four strains of C. albicans were resistant to FLC with a minimum inhibitory concentration (MIC) >64microg/mL and all were resistant to KTC and ITC (MIC>16microg/mL). The MIC for two strains of C. tropicalis were >16microg/mL for KTC and ITC, and >64microg/mL for FLC. It is worth highlighting that all of the strains tested were sensitive to AMB with the MIC varying from 0.25-1.0microg/mL. All strains of M. pachydermatis were sensitive to ITC with a minimum fungistatic concentration (MFC) 0.0075microg/mL. The MIC for 29 strains was the same (MFC0.0075microg/mL) for KTC. The MFCs for FLC varied from 1 to 16microg/mL, and for AMB, the MFC interval was 0.125-8microg/mL. There were no alterations in the classic phenotypic features of the strains of Candida spp. and M. pachydermatis isolated from dogs but, unlike M. pachydermatis, Candida spp. were much more resistant to azole antifungal agents.     

Genetic typing of Malassezia pachydermatis from different domestic animals

In the present study, random amplification of polymorphic DNA, which detects DNA polymorphism in fungal genomic DNA, was applied for genetic typing of Malassezia pachydermatis isolates. Fifty-five isolates from different domestic animals and body sites and the neotype strain CBS 1879 were characterized. Primers M13 and OPT-20 were used to analyse their genetic relatedness and similarity. This technique allowed us to distinguish four different genetic types. The predominant genetic type was observed in isolates recovered from different anatomical locations in all animals. It was the only genetic type found in cats, horse, goat and pig. The other three genetic types were observed only in isolates from external ear canals of dogs. Types II and IV were only recovered from external otitic ears and type III from healthy ears. An animal was colonised by more than one type of M. pachydermatis and different genetic types were detected in the same body site. Some genetic types were only isolated from diseased skin.     

Comparative analysis of the frequency, distribution and population sizes of yeasts associated with canine seborrheic dermatitis and healthy skin

The purpose of this study was to investigate the diversity of yeast associated with the degree of canine seborrheic dermatitis (SD) by anatomical sites. Fifty-seven samples were divided as 17 healthy skin, 20 with primary seborrheic dermatitis (PSD), and 20 with secondary seborrheic dermatitis (SSD). Yeast isolation and characterization were carried out based on microscopical features and biochemical properties. DNA analysis at the internal transcribed spacer I of 26S rDNA region was utilized for species confirmation. Four species of yeast consisting Malassezia pachydermatis, Malassezia furfur, Candida parapsilosis and Candida tropicalis recovered from examined dogs. M. pachydermatis and C. parapsilosis were isolated from all dogs, but C. tropicalis and M. furfur were recovered from 3 healthy dogs and one diseased dog, respectively. The number of M. pachydermatis and C. parapsilosis in diseased dogs was higher than that of healthy specimens (P<0.01). High frequency and population size of C. parapsilosis were closely associated to PSD, while those of M. pachydermatis were associated with both PSD and SSD (P<0.01). C. parapsilosis were predominant at the perianal area. This study demonstrated the co-colonization of M. pachydermatis and C. parapsilosis in large amounts and frequency associated with stage of disease and anatomical site.     

Fungal flora on cutaneous and mucosal surfaces of cats infected with feline immunodeficiency virus or feline leukemia virus

OBJECTIVE: To compare cutaneous and mucosal mycoflora in cats infected with FIV or FeLV with that in noninfected cats. ANIMALS: 85 client-owned cats; 24 seropositive for FIV, 10 seropositive for FeLV, 1 seropositive for both viruses, and 50 seronegative for both viruses. PROCEDURE: Cutaneous specimens were obtained from the coat and external acoustic meatus (ear canal) and mucosal specimens from the oropharynx and rectum. Fungi were isolated from specimens, using Sabouraud dextrose agar incubated at 27 or 37 C for cutaneous and mucosal specimens, respectively. RESULTS: Fungal colonies were cultured from at least 1 specimen from 83 of 85 (97.6%) cats. The most common fungal isolates were Aspergillus spp (cultured from 59.3% of all specimens), Penicillium spp (50.0%), Cladosporium spp (44.2%), Scopulariopsis spp (41.8%), and lipophilic yeasts of the genus Malassezia (31.4%). A greater diversity of fungal genera was isolated from retrovirus-infected cats, and Malassezia spp were more commonly recovered from these cats, compared with noninfected cats. Candida albicans, Cryptococcus neoformans, and dermatophytes (eg, Microsporum canis) were rarely isolated from any cat. Significant differences in frequency of isolation of C. neoformans and dermatophytes were not found between infected and noninfected cats. CONCLUSIONS AND CLINICAL RELEVANCE: Cats infected with FIV or FeLV may have a greater diversity of cutaneous and mucosal mycoflora than noninfected cats. However, infected cats may be no more likely than noninfected cats to expose humans to zoonotic fungi such as C. albicans, C. neoformans, and M. canis.     

Antifungal effect of Australian tea tree oil on Malassezia pachydermatis isolated from canines suffering from cutaneous skin disease

The lipophilic yeast Malassezia pachydermatis is part of the normal skin flora of most warm-blooded organisms. In a number of surveys it could be demonstrated that this yeast species might be involved in different skin diseases like seborrhoeic dermatitis, especially in dogs and cats. In order to look for an alternative therapeutic agent to the commonly used antimycotic and antiseptic synthetic substances the in vitro activity of Australian tea tree oil, the essential oil of Melaleuca alternifolia, against several strains of Malassezia pachydermatis was examined. All tested strains showed remarkably high susceptibility to tea tree oil. With these results the excellent antibacterial activity of tea tree oil is extended to a new group of fungal pathogens colonizing mainly mammals' skin. During the last ten years there was an increasing popularity of tea tree oil containing human health care products. The presented data open up new horizons for this essential oil as a promising alternative agent for topical use in veterinary medicine as well.     

Molecular heterogeneity in clinical isolates of Malassezia pachydermatis from dogs

Molecular investigation of 16 strains, conventionally identified to be Malassezia pachydermatis, isolated from dogs in Japan was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD band patterns of 13 clinical isolates were identical to that of standard strain of M. pachydermatis (CBS-1879). The other three clinical isolates were different from the standard strain of M. pachydermatis in RAPD patterns, and two of the three isolates were identical. About 620 bp genomic DNA fragments of the CHS2 gene were amplified from the same 16 clinical isolates of M. pachydermatis by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of the 16 clinical isolates revealed that the 13 strains were genetically very close to the standard strain of M. pachydermatis and the other two isolates were genetically close to the standard strain of M. furfur rather than M. pachydermatis. The remaining one isolate was phylogenetically distinct from all the seven Malassezia species reported so far.