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.     

Prevalence of Malassezia spp. yeasts in feline nail folds: a cytological and mycological study

Malassezia spp. yeasts are commensal organisms of mammal and avian skin, but little is known about their presence on the skin of healthy cats. The purposes of this study were to evaluate the prevalence of Malassezia spp. yeasts in feline nail folds and to identify the different species. Forty-six cats of different breeds were evaluated by cytological examination, and Malassezia spp. yeasts were seen in 61% of them. Yeasts were found in 100% of Devon Rex cats [mean 8.63/oil immersion field (high-power field - HPF)]. Conversely, only 42% of cats of other breeds (domestic short-haired and Persian) were positive (mean 0.59/HPF). Twenty-one cats of different breeds were subsequently evaluated by fungal culture. Malassezia pachydermatis was isolated from 52%, M. furfur from 38%, and M. sympodialis from 9.5% of the cats. More than one species was observed in eight of 21 cats, six of which were Devon Rex. Malassezia spp. yeasts are common inhabitants of feline nail folds, especially in Devon Rex cats, and the presence of a high number of yeasts on cytology correlates with the clinical observation of brown, greasy material in the nail folds. M. pachydermatis and two lipid-dependent species were isolated from both Devon Rex cats and cats of other breeds.     

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.     

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.     

Molecular characterization of Malassezia nana isolates from cats

Malassezia nana (M. nana) is a lipid-dependent yeast that has been isolated from cats and cows. Some sequence variability has been observed in the large subunit (LSU) and internal transcribed spacer (ITS) regions between strains isolated from cats and cows though these regions in M. nana isolates from cats alone have proven to be relatively conserved. In the present study, microsatellite PCR fingerprinting and β-tubulin gene sequence analysis were carried out on M. nana isolates from cats to investigate the genetic diversity of this species. Although a relatively small number of isolates were available, the similarity in the sequences of the β-tubulin and the microsatellite profiles indicate that a particular M. nana genotype colonizes cats. Moreover, all isolates obtained from animals with otitis externa had the same microsatellite fingerprinting pattern. Further studies of a wider population of M. nana isolates from other hosts and status disease are needed to establish that M. nana is a genetically homogeneous species. This is the first report of the characterization of the β-tubulin gene in Malassezia spp.     

Population sizes and frequency of Malassezia pachydermatis at skin and mucosal sites on healthy dogs

Skin and mucosal carriage of Malassezia pachydermatis was studied in 20 healthy pet dogs of various breeds and in 20 kennelled beagles. Using swabs, anal carriage was detected in 10 pet dogs and 11 beagles and the nose, mouth, prepuce and vulva were shown to be infrequently colonised. M pachydermatis was isolated from the external ear canal of 11 beagles and two pet dogs; both the population sizes and frequency of isolation were significantly (P<0·05) greater in the beagles. The yeast was infrequently isolated from the axilla and groin in low numbers using contact plates and detergent scrub samples but was often cultured from the lower lip and the dorsal interdigital spaces; isolation frequencies and population sizes in the two groups of dogs were not significantly different. These results demonstrate that the anus, external ear canal and lip and interdigital skin of healthy dogs are frequently colonised by M pachydermatis.     

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.     

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%).     

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.     

Malassezia species isolated from the intermammary and preputial fossa areas of horses

BACKGROUND: Malassezia-type yeasts previously have been observed on cytologic examination of the intermammary region of mares that presented with tail-head pruritus; topical antiyeast treatment resolved the pruritus. Further, Malassezia dermatitis has been observed in horses in intertriginous areas such as the udder and prepuce; the species of yeast was not confirmed. It is not known whether healthy mares or male horses can be carriers of this yeast in these body areas. HYPOTHESIS: Malassezia spp. are present in the intermammary region in healthy mares and the preputial fossa in healthy geldings. ANIMALS: Eleven healthy horses (5 mares and 6 geldings). METHODS: Samples of surface material were taken digitally from the intermammary area of 5 mares and the preputial fossa region of 6 geldings. The samples were examined cytologically and were cultured on modified Sabouraud's dextrose agar. The DNA from yeast colonies grown on the agar was extracted, and samples were assayed using fungal generic polymerase chain reaction (PCR) analysis. Amplicons with positive PCR results were sequenced and compared with sequences in the BLAST database search program. RESULTS: Of 44 attempts at culture, 5 yielded a species identified as Malassezia equi, and 2 yielded M slooffiae. In contrast, of 44 cytologic examinations, yeasts with the morphology of Malassezia spp. were seen in 40 samples. CONCLUSIONS AND CLINICAL IMPORTANCE: Due to its presence in healthy horses, finding of Malassezia-type yeast on cytologic examination may not incriminate it as a pathogen. Despite difficulty in culturing, cytologic examination was an effective tool to rapidly demonstrate the organism.     

Use of contact plates for the quantitative culture of Malassezia pachydermatis from canine skin

The Malassezia pachydermatis populations of the axilla and groin of 12 normal and 12 atopic dogs were compared using tape-strips and contact plates. When assessed by either method, the mean density of yeasts in the groin of the atopic dogs was significantly greater (P<0.05) than that of the normal dogs, suggesting that the cutaneous microenvironment of the groin region of the atopic dogs favoured colonisation by this yeast. Differences between the counts from the axilla were not significant. The frequency of isolation of yeasts from both dogs and sites was significantly higher (P<0.05 and P<0.001, respectively) in the atopic group. There was a very highly significant correlation (P<0.001) between the tape-strip counts and contact plate counts in the atopic group only. This study suggests that isolation of numerous M pachydermatis colonies from the axilla and groin of dogs using contact plates is indicative of elevated skin surface populations. The simplicity of the contact plate method makes it suitable for the routine quantitative culture of cutaneous M pachydermatis populations in dogs with dermatological disease.     

Comparison of two sampling techniques to assess quantity and distribution of Malassezia yeasts on the skin of Basset Hounds

Cytological examination using the tape-strip technique and fungal culture using contact plates with modified Dixon's medium were compared to evaluate the carriage of Malassezia yeasts on four cutaneous sites (left pinna, umbilical region, axilla and perianal area) in adult Basset Hounds. Twenty animals were included in the study. High numbers of Malassezia were isolated from at least one area in 100% of the animals. The frequencies of isolation and population sizes differed significantly according to anatomical location. They were greater on the pinna, followed by the umbilical area, axilla and perianal area. Fungal culture was more sensitive than cytology for the isolation of Malassezia yeasts. Frequencies of isolation were greater using this method, but population sizes were constantly smaller than with cytology.     

Malassezia pachydermatis isolated from normal and diseased external ear canals in dogs: a comparative analysis

To investigate the role of Malassezia pachydermatis as a pathogenic agent in canine otitis, a comparative analysis of isolates from normal and diseased external ear canals in dogs was undertaken. Specimens were collected from the ears of dogs with unilateral or bilateral otitis and from healthy dogs. Mycological analysis was by direct microscopy and fungal culture on Sabouraud's dextrose agar and Dixon's agar. Of the otitis specimens, 63.7% showed typical Malassezia cells on cytological examination. In samples taken from the healthy ears of dogs with unilateral otitis, only 21.43% (P<0.05) showed evidence of Malassezia. M. pachydermatis was identified cytologically and culturally in 57.53% (P<0.05), 14.29% and 30.0% of samples from the ears of dogs with otitis, from the healthy ears of dogs with unilateral otitis and from the ears of healthy dogs with no otitis. In the group with otitis associated with M. pachydermatis, the poodle was the most common breed (39.29%; P<0.05), whereas in the group without otitis, the German Shepherd breed was prominent (although this observation was not statistically significant). In both groups, the majority of dogs with M. pachydermatis were aged between 1 and 3 years (P<0.05). The higher incidence of M. pachydermatis isolated from the ears of dogs with otitis externa suggests a putative pathogenic role of this yeast in this condition.     

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.     

Identification and distribution of a novel Malassezia species yeast on normal equine skin

This study aimed to investigate the distribution of Malassezia species yeasts on the skin of healthy horses. Acetate tape samples were obtained from the lip, axilla, interbulbar region, groin and anus of 12 healthy horses. The samples were stained and examined microscopically and sites harbouring yeast-like organisms were identified. Contact plates were applied to the skin at these sites and cultured at 26 degrees C and 32 degrees C. No growth was obtained on horse blood, Sabouraud's dextrose or modified Dixon's agar. A pure growth of a Malassezia-type organism was obtained on Sabouraud's dextrose agar enriched with oleic acid when it was incubated at 30 degrees C. It was identified by 26S ribosomal DNA D1/D2 sequence analysis as a member of the genus Malassezia, and most closely related to Malassezia sympodialis. However, the level of sequence divergence indicated that it was a novel species.     

Attachment of Malassezia pachydermatis to the ear dermal cells in canine otitis externa.

To investigate the predominance of Malassezia pachydermatis (M. pachydermatis) as a causative agent of canine otitis externa, ear cerumen samples were observed for adhesion of M. pachydermatis to the cornified epithelial cells by light and electron microscopes. The yeasts appeared not to adhere to the cornified epithelial cells directly, but they seemed to exist in the proximity of the epithelial cells with an electron opaque halo-like space around them. Polysaccharide and lipid staining techniques were conducted to identify the substances existing in that space. Lipid substances, not saccharides, were observed around the yeasts and the cornified epithelial cells. These results suggested that in the canine ear canal malassezia yeast attachment to the cornified epithelial cells is mediated by lipids.     

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.     

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.     

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.