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.     

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.     

Comparative efficacies of oral ketoconazole and terbinafine for reducing Malassezia population sizes on the skin of Basset Hounds

The objective of this study was to compare the efficacy of oral ketoconazole and terbinafine for reducing population sizes of Malassezia yeasts on canine skin. Twenty-one Basset Hounds were randomised in three groups of seven according to Malassezia populations. Dogs in the first group were treated by oral administration of ketoconazole (Ketofungol) 200 mg, Janssen-Cilag) at 10 mg x kg-1, every 24 h with food, for 3 weeks. Dogs in the second group were treated by oral administration of terbinafine (Lamisil) 250 mg, Novartis) at 30 mg x kg-1, every 24 h with food, for 3 weeks. The seven remaining dogs were used as controls. Malassezia population sizes were assessed by use of contact plates on four cutaneous sites at days 7, 14 and 21. Both ketoconazole and terbinafine were effective in reducing the baseline levels of Malassezia organisms with no significant difference between the two drugs. In further studies, oral terbinafine should be evaluated for the management of canine cases of Malassezia dermatitis.     

Malassezia dermatitis in the dog: a retrospective histopathological and immunopathological study of 86 cases (1990–95)

A retrospective histopathological and immunopathological study was conducted on 86 dogs with Malassezia dermatitis. West Highland White terriers, English Setters, Shih Tzus, Basset Hounds, American Cocker Spaniels, spayed females, and castrated males were found to be at increased risk. The histopathological reaction pattern of lymphocytic superficial perivascular to interstitial dermatitis with parakeratotic hyperkeratosis, irregular epidermal hyperplasia, diffuse intercellular oedema and lymphocytic exocytosis was found to be consistent with a diagnosis of Malassezia dermatitis whether yeast were histologically visible (73.3% of the cases) or not (26.7%). Immunopathological studies revealed that 60– > 90% of the inflammatory cells within the epidermis, and 25–75% of those within the dermis were CD3+T lymphocytes, and that the only immunoglobulin-positive cells were dermal plasma cells.     

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.     

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%).     

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.     

Tuberculosis in five basset hounds

Five Basset Hounds (2 females and 3 males) under the age of 5 years, acquired systemic tuberculosis. We suspected tuberculosis in one dog, because it had histologic lesions similar to those in 4 dogs in which bacteria were identified as Mycobacterium avium complex. A review of canine tuberculosis revealed a similar diagnosis in a Basset Hound. The association of this infection in Basset Hounds suggests an inherited immunologic defect. Results of our survey suggest that the defect might exist in cell-mediated immunity.     

Use of etretinate for treatment of primary keratinization disorders (idiopathic seborrhea) in cocker spaniels, west highland white terriers, and basset hounds.

An open clinical trial was used to evaluate the synthetic retinoid, etretinate, for treatment of idiopathic seborrhea in Cocker Spaniels, West Highland White Terriers, and Basset Hounds. Clinical and histologic improvement was seen in the Cocker Spaniels. Etretinate had no beneficial effect on the skin disease of the West Highland White Terriers or the Basset Hounds. Etretinate treatment did not alter the type or degree of otitis externa. Clinical side effects were minimal. Relevant laboratory abnormalities were not detected.     

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.     

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.     

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.     

Colonisation status of Malassezia pachydermatis on the hair and in the hair follicle of healthy beagle dogs

Hair and hair follicle carriage of Malassezia pachydermatis was studied in 12 healthy beagle dogs. The yeast was isolated from hair clipped from the lip region at 13 sites in nine dogs but was less frequently recovered from the interdigital spaces on the forefeet and from two sites on the trunk. Population sizes at the lip were significantly greater (P < 0.01) than those at other sites. Skin biopsy specimens were obtained from the same sites and epidermal and follicular tissues dissected following immersion in 1 M CaBr(2). Epidermal carriage of M. pachydermatis was identified in nine biopsy specimens taken from five dogs. Hair follicle carriage was identified in five skin specimens (four foot, one lip) from three dogs. This study indicates that M. pachydermatis is readily recovered from the distal hair in healthy dogs and that hair follicle carriage is infrequent or that populations are low at that site.     

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.     

Factors associated with elevated cutaneous Malassezia p a chyderm a tis populations in dogs with Pruritic skin isease

The prevalences of breeds and concurrent diseases in a group of 40 dogs with pruritic skin disease associated with elevated cutaneous Malassezia pachydermatis populations were compared with samples of a dermatological hospital population. The ages and genders of the affected dogs were comparable to those of the dermatology population. Basset hounds, cocker spaniels and West Highland white terriers were significantly overrepresented. Concurrent diseases were diagnosed in 27 dogs, of which 15 were atopic. However, the prevalences of atopic disease, primary keratinisation defects and endocrinopathies in dogs with elevated cutaneous M pachydermatis populations were comparable to those in the dermatology population as a whole. These results indicate that certain breeds are predisposed to the development of elevated cutaneous M pachydermatis populations and that concurrent skin diseases can frequently be identified in affected dogs. However, the relationship between these concurrent diseases and abnormal M pachydermatis populations remains unclear.     

Comparison of orbital prosthesis motility following enucleation or evisceration with sclerotomy with or without a motility coupling post in dogs

Objective  To evaluate motility of silicone orbital implants and corneoscleral prostheses, with and without use of a motility coupling post (MCP) in dogs.
Animals studied  Eighteen mixed-breed dogs.
Procedures  The motility of an orbital silicone implant and corneoscleral prosthesis after enucleation ( n  = 6), evisceration ( n  = 6), or use of a MCP with evisceration ( n  = 6) in dogs were compared. One eye from each dog had surgery whereas the opposite eye was used as a control. Clinical evaluations were performed three times a week. Histopathology of the orbital tissues was performed 8 and 12 weeks after surgery.
Results  Implant motility in dogs with evisceration (vertical movement [VM] 8.04 ± 2.13; horizontal movement [HM] 11 ± 3.05) and evisceration with MCP (VM 9.61 ± 1.59); HM was significantly greater than the enucleation group (VM 0.51 ± 0.5; HM 1.22 ± 0.68) ( P <  0.01). Prosthetic motility in dogs with evisceration with MCP was significantly greater than in dogs with evisceration; dogs with evisceration had significantly greater motility than dogs with enucleation ( P <  0.01). No postoperative complications were observed in any of the groups. No significant abnormalities were noted on histopathology.
Conclusions  MCP placement in silicone orbital implants significantly enhanced the prosthetic motility in dogs. This study supports the use of MCP in silicone orbital implants to enhance corneoscleral prosthesis motility and cosmetics in dogs.     

P-80 Clinical aspects of chronic progressive pastern dermatitis in Belgian draft horses

The aim of this study was to evaluate the fungi present on the skin surface in healthy horses with a particular interest in Malassezia on predisposed skin areas. Twenty-five mares were included, each sampled on nine skin areas including those most likely contaminated by Malassezia : nostrils, lips, ear pinnae, base of the mane, pasterns, pectoral areas, lateral thorax, udder and perianal areas. Two sterile carpets were applied to the skin, and then used for inoculation of agar (Sabouraud's plus cycloheximide and chloramphenicol), both with and without lipids for isolation of different species of Malassezia . This method is currently successful in isolating Malassezia in dogs. Cultures were incubated at 32°C, observed on day 5 for Malassezia , and then maintained at 27°C for complementary morphological identifications. From 223 cultures on standard medium, only seven remained sterile. The most common fungi were filamentous (number of positive samples): Scopulariopsis (156), Penicillium (79), Acremonium (67), Aspergillus (53), Paecilomyces spp. (38), and Stachybotrys (33). Interestingly, Microsporum gypseum was also isolated (14 samples from seven areas) from 10 horses. Yeast or yeast-like fungi were Geotrichum (52), Trichosporon (21), and an additional 33 samples being positive for yeast other than Malassezia . Results from the other 223 cultures on lipid-rich agar were similar. Most of the fungi were isolated from the nine areas selected, but Malassezia was never isolated from 446 cultures.
Funding: Self-funded.     

Genotyping of Malassezia pachydermatis isolates from canine healthy skin and atopic dermatitis by internal spacer 1 (IGS1) region analysis

Isolates of Malassezia pachydermatis from healthy dog skin and from dogs with atopic dermatitis were molecularly characterized using internal spacer 1 (IGS1) region analyses, and their phospholipase A2 activity and pH growth profiles were then characterized in vitro. The percentage of isolates from healthy dogs that had the following IGS1 subtypes (isotype, %) were as follows: 1A, 6%; 1B, 27%; 1C, 11%; 2A, 6%; 2B, 6%; 3A, 11%; 3C, 3%; and 3D, 24%. In contrast, 9% of isolates from dogs with atopic dermatitis were isotype IB and 91% were isotype 3D, indicating that isolates of subtype 3D were the most prevalent in dogs with atopic dermatitis. Production of phospholipase A2 was statistically higher in isolates of subtype 3D than in the other subtypes. The subtype 3D isolates showed enhanced growth on alkaline medium compared with non-3D subtype isolates. The main clinical sign of canine Malassezia dermatitis is waxy exudates on the skin, which predispose the patient to development of a yeast overgrowth of the subtype 3D. Increased phospholipase A2 production may be involved in the inflammatory process associated with Malassezia dermatitis.     

Immunoglobulin G responses to Malassezia pachydermatis in healthy dogs and dogs with Malassezia dermatitis

Serum immunoglobulin G (IgG) responses of healthy dogs and dogs with Malasseziapachydermatis dermatitis were compared by Western immunoblotting. M pachydermatis CBS 1879 was disrupted mechanically and its proteins were separated and blotted on to nitrocellulose membranes before being incubated with sera from eight healthy beagles, eight Irish setters with gluten-sensitive enteropathy, 15 healthy basset hounds, and 30 dogs with Mpachydermatis-associated dermatitis, 20 of which were basset hounds. The mean (se) numbers of bands of immunoreactivity observed in the seborrhoeic basset hounds (10.7 [0.4]) and affected mixed-breed dogs (9.4 [0.9]) were significantly greater than in the beagles (3-0 [1.0]), Irish setters (5.5 [1.1]) and healthy basset hounds (5.6 [0.7]). The number of bands identified was correlated (r(s) = 0.76, P < 0.001) with the anti-M pachydermatis IgG values measured by ELISA in a previous study. Most of the dogs were immunoreactive towards the 132, 66 and 50 to 54 kDa proteins and the affected dogs were also usually reactive towards the 219, 110, 71 and 42 kDa proteins.     

Collection of peripheral blood CD34+ progenitor cells from healthy dogs and dogs diagnosed with lymphoproliferative diseases using a Baxter-Fenwal CS-3000 Plus blood cell separator

Background

Canine peripheral blood mononuclear cell (PBMC) apheresis using a Baxter‐Fenwal CS‐3000 Plus automated blood cell separator has not been reported.

Objective

To determine the feasibility and safety of using a CS‐3000 Plus blood cell separator with a small volume separation container holder (SVSCH) and small volume collection chamber (SVCC) to harvest canine PBMCs from dogs weighing <50 kg.

Animals

Eight healthy mongrel dogs and 11 client‐owned dogs in clinical remission for lymphoproliferative diseases (LPD).

Methods

In this prospective study, aphereses were performed using a Baxter‐Fenwal CS‐3000 Plus blood cell separator, with or without recombinant human granulocyte colony‐stimulating factor (rhG‐CSF) treatment.

Results

Aphereses from 6 healthy dogs given rhG‐CSF yielded an average of 1.1 × 10⁷ ± 8.2 × 10⁶ CD34+ cells/kg. Aphereses from LPD dogs given rhG‐CSF yielded an average of 5.4 × 10⁶ ± 3.25 × 10⁶ CD34+ cells/kg (P = .17). Higher hematocrit in both groups of dogs receiving rhG‐CSF correlated with an increased number of CD34+ cells/kg harvested (healthy, P = .04; LPD, P = .05). Apheresis was well tolerated by all dogs.

Conclusions and Clinical Importance

Canine PBMC apheresis using the Baxter‐Fenwal CS‐3000 Plus cell separator with an SVSCH and SVCC is a feasible and safe option for harvesting an adequate number of CD34+ peripheral blood progenitor cells from dogs weighing ≥17 kg for hematopoietic cell transplantation.