Poster Abstracts Topic 1: SKIN BIOLOGY

Veterinary Dermatology 2004, 15 (Suppl. 1), 41 69 Topic 1: SKIN BIOLOGY P-1 Evaluation of the influence of sex, diet and time on hair coat characteristics of cats P. BOURDEAU, K. W. TAYLOR, P. NGUYEN and V. BIOURGE National Veterinary School of Nantes, Nantes, France; Royal Canin, Aimargues, France Skin physiology in cats has received little attention. The aim of this study was to evaluate the long-term influence of sex, time and the level of dietary fat and energy on the dynamics and qualities of the hair coat. Twenty-four European short-haired laboratory cats were followed over a 1-year period. They were divided into eight groups of three, according to: sex (12 males and 12 females), sexual status (intact or neutered) and diets [(high energy 4300 kcal/kg as fed, 21% fat) vs. (moderate energy 3500 kcal/kg as fed, 10% fat)]. Both diets were fed for 6 months to all cats following a cross-over design. The following parameters were evaluated throughout the study: thickness of hair coat and hair lengths (neck, rump, lateral, flank), hair regrowth (after periodic clippings of 25 cm 2 areas), and telogen/anagen ratio. The thickness of the hair coat initially varied from 1.2 1.7 cm on the neck, 1 1.4 cm on the rump, 1.8 2.5 cm on the flank, and hair shaft lengths were 1.7 2.5, 3.7 3.9 and 2.5 3.2 cm, respectively. Comparison of values revealed few statistical differences: increase of the thickness of hair coat in neutered cats (male and female) during the study, and increase of the length of lateral hairs in all groups during the study. Over all periods and in all groups, the curve of growth was similar (rapid then slower). Some transient variations were attributed to temporary changes in ambient conditions. In conclusion, neither sex, nutrition or season (in housed cats) influenced the general quality of hair coat, in particular hair regrowth. Funding: Royal Canin. P-2 Evaluation of the influence of sex, diet and time on skin ph and surface lipids of cats P. BOURDEAU, K. W. TAYLOR, P. NGUYEN and V. BIOURGE National Veterinary School of Nantes, Nantes, France; Royal Canin, Aimargues, France Skin lipids and ph are two factors classically considered of importance in homeostatic characteristics of skin. Skin physiology in cats has received little attention. The aim of this study was to evaluate the long-term influence of sex, sexual status, season, and dietary fat and energy on these parameters. Twenty-four European short-haired laboratory cats, 14 months of age, were followed over a 1-year period. They were divided into 8 groups of three, according to: sex (12 males and 12 females), sexual status (intact or neutered) and diets [(high energy 4300 kcal/kg as fed, 21% fat) vs. (moderate energy 3500 kcal/kg as fed, 10% fat)]. Both diets were fed to all cats for 6 months following a cross-over design. Parameters regularly evaluated were skin ph and hair total lipid content (extraction from samples of 0.6 1.2 g of clipped hairs). The ph of the skin varied from 6.6 6.8 initially to 7.2 7.4 at the end of the study. This increase was significant only in intact animals (male and female). The dietary changes did not affect skin ph. Hair total lipid content was not affected by sex or the diets but slightly increased in all groups over the study period from 1.5 2.4 to 2.4 3.3%. In conclusion, skin ph appeared to be potentially modified by sexual status, but not by sex or dietary lipids and energy. Hair lipids do not appear to be affected by sex or increases in dietary lipids and energy. The slight increase in ph and hair lipids during the study was attributed to the change from young adult to adult stage of all cats. Funding: Royal Canin. Topic 2: BACTERIAL DISEASES P-3 Involvement of the accessory gene regulatory (agr) system in quorum sensing by Staphylococcus intermedius J. M. L. SUNG, P. D. CHANTLER and D. H. LLOYD Royal Veterinary College, University of London, Hatfield, Herts, UK Toxic exoproteins are believed to be involved in the pathogenesis of Staphylococcus intermedius pyoderma and bacterial overgrowth, although these diseases are poorly understood. This study aims to determine whether quorum sensing, a phenomenon in which cells express particular characteristics only when population densities exceed certain levels, might be involved. Production of virulence factors by S. intermedius was hypothesized to be initiated by the agr system via signalling molecules generated as cell density increased, as in S. aureus. Initial studies (agrspecific PCR, genome walking) confirmed the presence of sequences in S. intermedius related to the agr of S. aureus. To test the hypothesis that expression of RNAIII (effector of the agr system) and two exotoxins, leukotoxin and enterotoxin C, are initiated by an environmental signal generated during bacterial growth, experiments were performed in which spent culture supernatants from different phases of growth were added to early exponential cultures of S. intermedius in which the genes are minimally expressed. Through real-time quantitative PCR, it was shown that concentrated supernatant of midexponential and postexponential cultures from S. intermedius strains 3 and 27 up-regulated expression of genes encoding for RNAIII, leukotoxin and enterotoxin in strain 3. This indicates that the activating substance accumulates in supernatant during growth. In contrast, S. intermedius was insensitive to spent medium from S. aureus strain J3964. This suggests that S. intermedius uses quorum sensing to monitor the proximity of other signal-producing cells and thereby adjusts its expression of virulence genes appropriately to facilitate species-specific and intraspecies cell-cell communication. Funding: Nestle Purina Research Company. Ó 2004 ESVD and ACVD 41

42 P-4 Canine recurrent pyoderma: a multicenter prospective study E. BENSIGNOR and P. A. GERMAIN Veterinary Referral Service, Paris, France; Cabinet de Dermatologie Ve te rinaire, Me rignac, France Bacterial pyoderma is one of the most frequent skin diseases in dogs. Recurrent pyoderma is often secondary to an underlying skin disease, but no epidemiological study has been published on the subject to the authors knowledge. This study was designed to prospectively evaluate the causes responsible for recurrent pyoderma in dogs. Dogs presenting with a history of more than three episodes of skin infection in the last year were included in the study. For each case, epidemiological and clinical data were collected. Pyoderma was confirmed by the clinical signs, the demonstration of bacteria on microscopic examination of cytological smears and a positive culture. Each animal was treated with an appropriate course of antibiotics until resolution of signs of pyoderma. Depending upon the presence of pruritus, appropriate diagnostic tests were performed: skin scrapings, acaricidal trial, flea treatment, elimination diet, intradermal testing, biopsies, endocrinological tests, leishmaniasis and ehrlichiosis serology, antinuclear antibody testing. Thirty dogs (14 males and 16 females) of 19 different breeds, aged from 1 to 12 years (mean 4.9 years) were included. Diagnosis was folliculitis (44%), folliculitis and furunculosis (20%), furunculosis (20%), cellulitis (10%). Staphylococcus intermedius was isolated in 97% of cases. The following underlying diseases were identified: atopic dermatitis (60%), food allergy (7%), flea allergy (7%), hypothyroidism (7%), hyperestrogenism (4%), demodicosis (4%), and zinc-responsive dermatosis (4%). In two dogs, no underlying cause could be identified. Atopic dermatitis is the most common disease associated with recurrent pyoderma in dogs. Funding: Pfizer Animal Health. P-5 Importance of psychogenic factors in canine recurrent pyoderma M. NAGATA and K. SHIBATA Animal Dermatology Center, ASC, Chofu, Tokyo, Japan Recurrent pyoderma is a relapsing bacterial infection of the skin. Evaluation of the primary predisposing factors is essential, but they are not always easy to determine. We reported that psychogenic factors should be considered as one cause of canine dermatoses featuring pruritic behavior such as grooming and scratching, and predicted that it could be added as a cause of recurrent pyoderma. The purpose of this study was to evaluate the roles of psychogenic factors in canine recurrent pyoderma. This study examined 21 dogs with recurrent pyoderma, which showed relapsing but antibiotic-responsive skin lesions. Predisposing factors, especially allergy, infection, endocrine disorders, congenital factors and improper treatments, were carefully ruled out with standard diagnostic procedures. Diagnosis of psychogenic factors was based on fulfillment of the following features: (1) distinctive area of broken hairs or asymmetric erythema, (2) incidental pruritic behavior related to emotionally unstable situation or physiological intervention, (3) existence of environmental antecedent triggers or concurrent psychiatric symptoms, and (4) requirement for psychogenic treatment, consisting of behavior modification and psychopharmacological therapy in some. The incidence of psychogenic factors and the efficacy of the treatment in recurrent pyoderma were evaluated. In these 21 dogs, 10 cases (47.6%) were compatible with the presence of psychogenic factors, and seven cases (33.3%) were improved with behavioral treatment. Psychogenic factors should be considered in the aetiology of so-called idiopathic canine recurrent pyoderma. Further investigation is needed to understand the relationship between skin barrier dysfunction and psychogenic factors. P-6 Double-blinded comparative study of the efficacy of azithromycin and cephalexin in canine pyoderma A. D. BLANCO and A. WOLBERG College of Veterinary Medicine, University of Buenos Aires, Capital Federal, Argentina A double-blinded randomized, controlled study was carried out with the objective to evaluate the comparative efficacy of two antibiotics (azithromycin and cephalexin) in dogs with superficial pyoderma. Thirtyseven dogs with clinical signs of superficial pyoderma were included in the study. The diagnosis of superficial pyoderma was based on history, clinical signs and cytology. The presence of underlying diseases was investigated through complementary diagnostic methods after clinical resolution of the pyoderma. Animals were randomly assigned to one of two treatment groups: cephalexin 30 mg/kg twice daily for 21 days, or azithromycin 20 mg/kg twice daily for 21 days. Response to therapy was considered to be satisfactory if complete recovery was achieved by day 14 of treatment. The effects of the two treatments were compared using the chi-square test. Twenty dogs received cephalexin and 17 received azithromycin. Nineteen of 20 dogs treated with cephalexin achieved complete recovery on day 14, with no side effects reported. Ten of 17 dogs treated with azithromycin achieved complete recovery on day 14, but therapy was discontinued in one dog because of vomiting. Cephalexin was significantly more effective than azithromycin (P = 0.0077). Azithromycin may be a therapeutic alternative in those patients in which cephalexin cannot be used.

43 P-7 Treatment of canine acne with doxycycline: an open trial E. BENSIGNOR and E. GUAGUERE Veterinary Dermatology Referral Service, Paris, France; Veterinary Dermatology Referral Service, Lomme, France The purpose of this study was to determine whether doxycycline (Ronaxan) decreased the severity of localized lesions of canine acne. Dogs were enrolled if they exhibited skin lesions localized to the chin compatible with canine acne, and if cytological smears showed a bacterial invasion and bacteriological samples grew Staphylococcus intermedius colonies. Demodicosis and dermatophytosis were excluded by skin scrapings and fungal culture, respectively. Lesions (erythema, lichenification, papules, pustules and excoriations) were graded with a quantitative 4-point scale (maximal total score 15). Extent of lesion was also estimated. Pruritus was graded on a 4-point scale. Scores were given at the time of inclusion and at day 30 after treatment with doxycycline 10 mg/kg once daily. Twelve dogs (10 males and two females) representing seven breeds were included. Good improvement of both lesional (9.4 before treatment and 2.3 after treatment) and extent scores (43% before treatment and 8% after treatment) was achieved. Pruritus was also markedly improved from 2.08 before treatment to 0.42 after treatment. One dog was not significantly improved after 1 month. Results of this trial suggest that doxycyline is useful for reducing the severity of localized skin lesions of canine acne. P-8 Antibacterial efficacy of 0.1% chitosan solution against Staphylococcus intermedius in dogs H. H. JEONG, J. Y. KIM, K. W. LEE and T. H. OH College of Veterinary Medicine, Kyungpook National University, Daegu, Korea Antibacterial efficacy of 0.1% chitosan solution against Staphylococcus intermedius in dogs was determined by the modified detergent scrub quantitative technique in 10 adult mongrel dogs. For comparison, other antibacterial agents including 2.5% benzoyl peroxide, 0.5% chlorhexidine acetate, 0.1% chitosan solution combined with 2.5% benzoyl peroxide, and chitosan combined with 0.5% chlorhexidine were used. The antibacterial activity of chitosan was inversely affected by ph with higher activity at lower ph value and exerted the highest activity at ph 4.5 in vitro. In nontreated bacteria-challenged control sites, S. intermedius was found to have counts of 6.95 ± 0.53 cfu/cm 2 of skin. The counts of recovered Staphylococcus intermedius (cfu/cm 2 ) from the treated sites were 3.25 ± 0.98 for 0.1% chitosan solution, 0.68 ± 1.13 for benzoyl peroxide, 3.14 ± 0.55 for chlorhexidine, 0.48 ± 0.56 for chitosan solution combined with benzoyl peroxide, and 2.55 ± 0.88 for chitosan solution combined with chlorhexidine. Each S. intermedius recovery value from the five treated sites was significantly (P < 0.01) lower than that from the nontreated bacterial-challenged control site. The 0.1% chitosan solution was efficacious against Staphylococcus intermedius in dogs. However, the combinations of chitosan with benzoyl peroxide and with chlorhexidine were shown to exert higher activity when compared to those of chitosan alone and benzoyl peroxide or chlorhexidine alone. Thus, 0.1% chitosan solution was considered to have antibacterial activity against Staphylococcus intermedius and have potential for clinical applications in the treatment of pyoderma in dogs. P-9 CD8 and CD4 lymphocyte populations in blood from healthy dogs and dogs naturally infected with Ehrlichia canis C. LORENTE-ME NDEZ, A. SAINZ and M. A. TESOURO Faculty of Veterinary Medicine, University Cardenal Herrera CEU, Valencia, Spain; University Complutense, Madrid, Spain; University of Leo n, Leo n, Spain The purpose of this study was to assess the presence of an immune dysregulation in dogs naturally infected with Ehrlichia canis. We compared CD4 and CD8 lymphocyte populations in a group of dogs with ehrlichiosis and a group of healthy dogs. Thirty-seven dogs with natural canine ehrlichiosis (Group A) and 15 unaffected dogs (Group B) were included in the study. The study of lymphocyte populations was made by flow cytometry. Monoclonal antibodies against CD3, CD4 and CD8 were used. Results obtained in this study show an inversion in the CD4:CD8 ratio in the blood of dogs with ehrlichiosis when compared with the CD4:CD8 ratio in blood from control dogs. In healthy dogs, the number of CD4 T lymphocytes was greater than CD8 T lymphocytes. However, in dogs with ehrlichiosis, the number of CD8 T lymphocytes was greater than CD4 T lymphocytes. The percentage of CD4 T lymphocytes was lower in dogs with ehrlichiosis than in healthy dogs, but significant differences were not found between both groups. However, mean value of the percentage of CD8 T lymphocytes (with respect to lymphocyte population) in dogs with ehrlichiosis (45.81 ± 20.90%) was significantly higher (P =0.001) than in healthy dogs (25.41 ± 13.63%). These results confirm the presence of an alteration in the T lymphocyte subpopulations of dogs with ehrlichiosis, suggesting the presence of an immune dysregulation.

44 Topic 3: FUNGAL DISEASES P-10 Comparison of carpet and toothbrush methods for the detection of asymptomatic carriage of dermatophytes in cats P. BOURDEAU, F. COSTIOU and C. PERON National Veterinary School of Nantes, Nantes, France Diagnosis of Microsporum canis infection is a challenge in cats with suspected asymptomatic carriage. The aim of this study was to compare the carpet method and the toothbrush method of sample collection for dermatophyte culture. The study was conducted on apparently healthy cats in a chronically infected cattery. Sampling was performed with both a sterilized piece of carpet and a toothbrush applied to the entire body surface. Samples were inoculated on Sabouraud s media (with chloramphenicol and cycloheximide). Multiple (6) applications with the toothbrush were necessary to obtain a similar surface area of inoculation. Cultures were incubated at 27 C, then examined from day 5 to 21 for the presence and number of colonies of M. canis and other fungi. Some samples were inoculated twice (initially negative or with a low number of M.canis). A total of 112 cultures were performed (14 duplicates) from 44 cats. On day 21, infection was detected in 23 cats (54.7%): 20 (87%) with the carpet method and 21 (91%) with the toothbrush method. None of the replicated cultures allowed the detection of new cases but three were subsequently negative. In eight cases, only one to six colonies of M. canis were obtained. Both methods used are easy to perform on cats and have a similar sensitivity of approximately 90%. The carpet method is, however, much less expensive, easier to prepare, mail, store and inoculate. The lack of growth in some duplicate cultures (initial development of one or two colonies) may explain the limits of diagnosis of infection and justify the need for multiple inoculations of agar with the toothbrush method. P-11 Occurrence of dermatophytes in dogs and cats and their susceptibility to antifungal drugs I. ZDOVC, J. MICˇUNOVI, T. PIRSˇ and M. OCEPEK Veterinary Faculty, University of Ljubljana, Ljubljana, Slovenia The purpose of this study was to evaluate the frequency of dermatophytosis in dogs and cats with skin disorders and in clinically healthy animals suspected to be a source of human dermatophyte infection. Over a 2-year period, 776 samples from 339 dogs and 437 cats were examined. Samples were cultured on Sabouraud s dextrose agar (with chloramphenicol and actidione) for 2 3 weeks. Antifungal susceptibility tests for ketoconazole, fluconazole, itraconazole, amphotericin B and flucytosine (E test) were performed to establish the MIC values. Different dermatophyte species from the genus Microsporum and Trichophyton (M. canis, M. gypseum, T. mentagrophytes, M. persicolor) were isolated from 194 samples (25.0%). Among 57 positive dogs, M. canis was isolated from 26 (45.6%), T. mentagrophytes from 28 (49.1%), M. gypseum from two (3.5%) and M. persicolor from one (1.7%). Among 137 positive cats, M. canis was isolated from 124 (90.5%), T. mentagrophytes from 12 (8.7%) and M. gypseum from one (0.7%). These results are different to those reported in the past, where M. canis was the predominant dermatophyte species in both dogs and cats. In the last 2 years, the occurrence of T. mentagrophytes infections in carnivores has evidently increased, especially in dogs. One of the possible reasons is the increasing number of rabbits and rodents being kept as pet animals. As T. mentagrophytes is often isolated from these animals, they may serve as a source of infection. The results of susceptibility testing to antifungal agents were similar to those obtained in previous testing with most dermatophytes susceptible to ketoconazole and itraconazole, but resistant to fluconazole. P-12 Comparative study of griseofulvin and terbinafine therapy in the treatment of canine and feline dermatophytosis A. C. BALDA, M. OTSUKA, W. GAMBALE and C. E. LARSSON Veterinary Teaching Hospital, University of Sa o Paulo, Sa o Paulo, Brazil; Institute of Biomedical Sciences, University of Sa o Paulo, Sa o Paulo, Brazil In domestic carnivores, dermatophytosis is caused mainly by two fungal genera: Microsporum spp. and Trichophyton spp. These diseases are anthropozoonoses of significance in public health. The goal of this study was to compare the efficacy of griseofulvin and terbinafine in the therapy of the dermatophytosis. Thirty-five animals were included in three protocols of treatment. Griseofulvin was used at a dosage of 50 mg/kg once daily. It was effective in 100% of the cases, with no side effects, and the average time until cure was 41 days. Terbinafine was used at a dosage of 5 mg/kg once daily. It was effective in 81.3% of cases, no side effects were observed, and the average time until cure was 21 days. Terbinafine used at a dosage of 20 mg/kg once daily was similarly effective. However, at this higher dosage, the average time to cure was 33 days and side effects such as vomiting, diarrhoea and an increase in transaminase and alkaline phosphatase serum activity were seen in 16.6% of the animals. This study demonstrated that terbinafine is a good therapeutic alternative, although griseofulvin is still the first-choice drug for the treatment of dermatophytosis in dogs and cats.

45 P-13 Dermatophytosis in domestic cats: treatment with lufenuron R. R. RAMADINHA, R. REIS, S. G. CAMPOS, S. S. RIBEIRO and P. V. PEIXOTO Veterinary Institute, Federal Rural University of Rio de Janeiro, Rio de Janeiro, Brazil Preliminary studies showed that lufenuron inhibits chitin synthesis, a dermatophyte cell wall constituent, and may be effective in the treatment of dermatophytosis. Our purpose was to evaluate the efficacy of lufenuron in the treatment of feline dermatophytosis. Forty-six cats (Persians and mixed-breed cats from 1-month to 4-years old) naturally infected with Microsporum canis were included in this study. Fifteen cats were treated isolated in cages in the veterinary hospital and 31 were treated in their home environment (some with access to the outdoors). Dermatophyte skin lesions were seen in 29 animals while 17 other cats were asymptomatic carriers. Wood s lamp, direct microscopic examination of hairs, fungal culture and skin biopsies were used for the diagnosis. Affected cats and all in-contact animals received lufenuron at a dose of 120 mg/kg every 21 days for four treatments. Of the 29 symptomatic cats treated with lufenuron, 70% recovered within 21 days and 28% within 42 days of initiation of therapy. One cat had only partial recovery and another was euthanized. Negative fungal culture was recorded only after the fourth dose of lufenuron in 98% of affected cats and 100% of asymptomatic carriers. There was no difference in clinical response to lufenuron between the cats treated in their home environment and those treated in the veterinary hospital. Side effects were not observed, thus the drug proved to be safe and effective for the treatment of dermatophytosis. Funding: Novartis. P-14 Use of lufenuron in the therapy of canine and feline dermatophytosis in São Paulo (Brazil) M. SONODA, A. C. BALDA, M. OTSUKA, W. GAMBALE and C. E. LARSSON Faculdade de Medicina Veterina ria, Universidade de Sa o Paulo, Sa o Paulo, Brazil; Instituto de Cieˆncias Biome dicas, Universidade de Sa o Paulo, Sa o Paulo, Brazil The goal of this study was to evaluate the efficacy of lufenuron in the treatment of animals with dermatophytosis [M. canis or M. gypseum (n =48) and T. rubrum (n =1)]. The study was performed over a 2-year period. The 49 animals (23 dogs and 26 cats) were randomly separated into four groups, each one receiving 50 120 mg/kg of lufenuron, divided into one to four doses administered at various intervals. Liver enzyme measurement were performed on all animals. The results showed clinical and mycological cure in 22 60% of the animals treated in the study, with the highest efficacy in the group treated with 120 mg/kg given in two doses at a 3-week interval (group IV). No systemic or cutaneous side effects or changes in liver enzyme levels were observed in any of the animals. Persian cats and Yorkshire terrier dogs had the highest rates of therapy failure. Lufenuron was more effective in dogs than cats. Morphological changes occurred in isolated Microsporum spp. colonies on culture. P-15 Identification of Malassezia spp. isolated from a cat and cattle, including a new species, M. nana A. HIRAI, R. KANO, K. MAKIMURA, H. YAMAGUCHI and A. HASEGAWA School of Veterinary Medicine, Nihon University, Kameino, Fujisawa, Kanagawa, Japan; Institute of Medical Mycology, Teikyo University, Otsuka, Hachioji, Tokyo, Japan Isolates from a cat and cattle with or without otitis externa in Japan and Brazil were thought to be Malassezia spp., but were clearly distinct from the other nine species of Malassezia which have been reported by molecular analysis, including M. dermatis and M. japonica. These isolates resemble M. dermatis and M. sympodialis morphologically and physiologically, but can be distinguished from these two species by their ability to use cremophor EL as the sole source of lipid and to split esculin. These isolates were classified as a novel species, Malassezia nana, by mycological examination as well as molecular analysis. Methods to identify seven Malassezia species have been reported, but they do not include the three new species, M. nana, M. dermatis and M. japonica. In this study, we described a practical approach to the identification of all 10 species of Malassezia yeasts from clinical material. We improved the identification system by examining lipid requirement, catalase reaction, precipitate production on modified Dixon agar, utility of cremophor EL, ability to split esculin and the shape of cells. This approach represents an identification scheme for Malassezia spp. without elaborate equipment. P-16 Treatment of Malassezia otitis in dogs: a comparative field trial E. BENSIGNOR Dermatology Referral Service, Paris, France Otitis externa (OE) is a very common disease of dogs. Various treatments are available for OE, but most commercial preparations contain a mixture of antifungal, antiinflammatory drugs and antibiotic agents. The aim of this study was to evaluate if a commercial product containing clotrimazole, marbofloxacin and dexamethasone (Aurizon Ò ) was more effective than the use of a topical antifungal agent econazole (Daktarin Ò ) used alone to treat Malassezia otitis. Dogs presenting with bilateral OE and numerous yeasts on cytological examination were included. Twenty dogs aged from 1 to 14 years old were treated in this trial (40 ears, 20 in each group). Each case was treated with Aurizon Ò (four drops) in one ear and Daktarin Ò in the other ear. Criteria analysed were pruritus (visual analogue scale), erythema, amount of cerumen, and the number of yeasts on cytologic smears. All animals demonstrated a good improvement in Malassezia counts after either treatment. Significant differences were noted in favour of Aurizon Ò for erythema, cerumen, and pruritus on various days of evaluation. This study demonstrated the superiority of a mixture of corticosteroids and antifungal agent in comparison with an antifungal agent used alone for the treatment of Malassezia OE.

46 P-17 In vitro activity of posaconazole and other antifungals against Malassezia pachydermatis isolated from dogs P.BOURDEAU,A.M.MARCHANDandF.ETORÉ National Veterinary School of Nantes, Nantes, France; Schering-Plough Ve te rinaire, Levallois-Perret, France The aim of this study was to determine the in vitro antifungal activity of several antifungal drugs (posaconazole, nystatin, miconazole and clotrimazole) against Malassezia pachydermatis with microdilution and agar dilution techniques. Malassezia pachydermatis isolates were obtained from the skin and ears of dogs. Tests on solid media were performed using 25-well Petri dishes (2 ml/well containing Sabouraud s dextrose agar and diluted antifungal drug) inoculated with 5 ll suspensions of M. pachydermatis. Microtitre broth dilution used 96- well microtitre plates containing Sabourauds dextrose broth and appropriate dilutions of antifungal drugs, inoculated with 10 ll standard suspensions of M. pachydermatis. Plates were inoculated in duplicate and incubated at 30 C for 5 days and growth assessed. The four antifungal drugs were tested in 10 dilutions (4.0-0.007 lg/ ml for posaconazole, and 32 0.06 lg/ml for clotrimazole, miconazole and nystatin). Results obtained for 83 strains of M. pachydermatis and a control reference strain (CBS 1879) exhibited the same pattern. Results of the MIC between microtitre and agar methodologies showed no significant differences ( 2-fold) across all drugs. For both solid and liquid methods, posaconazole was the most effective antifungal drug of the four tested with MIC 90 of 1 2 lg/ml for posaconazole, 16 32 lg/ml for clotrimazole, and 32 lg/ml for miconazole and nystatin. Funding: Schering-Plough. P-19 Sporotrichosis in São Paulo (Brazil): clinical and epidemiological features M. OTSUKA, R. C. C. CASTRO, N. S. MICHALANY, R. LUCAS, C. E. LARSSON Jr and C. E. LARSSON Faculdade de Medicina Veterina ria, Universidade de Sa o Paulo, Sa o Paulo, Brazil; Escola Paulista de Medicina, Universidade Federal do Estado de Sa o Paulo, Sa o Paulo, Brazil Human and animal sporotrichosis is a relatively common zoonotic disease in the southwest region of Brazil. In a recent outbreak (1999 2000) involving 66 humans in Rio de Janeiro, 79% of patients reported previous contact with infected cats. Over a 17-year period (1986 2002), 25 cases of feline sporotrichosis were diagnosed at our hospital (1.5 cases/year). Among these cases, 80% were male and had the cutaneous form of the disease, and 21 cases (84%) were in mixed-breed animals. The diagnosis was established based on histopathology (H&E and PAS stain) as well as fungal culture (95%) and cytologic examination (82.5%) of the cases. In the 25 homes of these animals, 15 (60%) were sporotrichosis cases of in-contact animals. Humans were infected in seven (28.5%) of the homes. Eighteen of the 49 dogs and cats (36.7%) in homes with infected humans were also affected. Itraconazole at 10 mg/kg once daily was successful even in FIV and FeLV positive cats. P-18 Cryptococcosis in São Paulo (Brazil): clinical and epidemiological features (1992 2003) S.C.CHIESA,R.C.CASTRO,M.OTSUKA,N.S. MICHALANY, C. E. LARSSON Jr and C. E. LARSSON Faculdade de Medicina Veterina ria, Universidade de Sa o Paulo, Sa o Paulo, Brazil; Escola Paulista de Medicina, Universidade Federal do Estado de Sa o Paulo, Sa o Paulo, Brazil Cryptococcosis is a relatively frequent disease in the southwest region of Brazil. We have diagnosed 19 cases of cryptococcosis (1.6 cases/year) at our hospital in the last 12 years. Cats were most frequently affected (18 feline and one canine) among our cases. The disease was most frequent in purebred Siamese cats (55.5 %), and in male cats (77.8 %) with an age predisposition of 4- to 6- year-old animals (56.2%) and 7- to 9-year-old animals (18.8 %). The diagnosis was established by cytologic and histopathologic examination, fungal culture and serologic evaluation. All cases was caused by Cryptococcus neoformans serotype AD. We used fungal culture as the gold standard for the diagnosis. Histopathology was positive in 93.3% of cases and the cytologic examination in 75% of cases. Most cats were FIV and FeLV negative. Ocular involvement was found only in the canine case which is the first report of canine cryptococcosis in Brazil. Itraconazole (10 mg/kg once daily) as the sole therapy or in conjunction with 5-fluorocytosine was effective in 95% of the patients.

47 P-20 Clinical, cytopathological and histopathological evaluation of sporotrichosis in experimentally infected cats M. R. FARIAS, S. R. V. S. FRANCO and V. E. FABRIS School of Veterinary Medicine, Pontifıćia Universidade Cato lica do Parana Sa o Jose dos Pinhais, Parana, Brazil; School of Veterinary Medicine and Zootechnics, Universidade Estadual Paulista, Botucatu, Sa o Paulo, Brazil; Medical School, Universidade Estadual Paulista, Botucatu, Sa o Paulo, Brazil The purpose of this study was to evaluate the clinicopathological aspects of experimental sporotrichosis in cats and compare the sensitivity of cytopathology, histopathology and culture as diagnostic tools in different phases of the infection. Twenty adult, mixed-breed cats (10 males and 10 females) were inoculated subcutaneously with 10 6 fungal microorganisms. Clinical examination was performed weekly. Cytopathologic, histopathologic and culture examinations were performed at 15, 30 and 60 days postinoculation. Culture of multiple organs was performed after euthanasia at 30 (10 cats) and 60 (10 cats) days postinoculation. Friedman parametric and nonparametric statistical analysis were applied to the results. The nodular, tumoral and necrotic lesions progressed significantly until day 30 postinoculation, and partial spontaneous regression occurred at day 60, particularly in males. An intense inflammatory pyogranulatomous and lymphocytic infiltrate with rare giant cells and sparse fibrosis associated with numerous, pleomorphic, intra- and extracellular fungal cells were observed on day 30. These findings gradually decreased by day 60. Despite the inflammatory granuloma associated with feline sporotrichosis, a tendency for dissemination was observed, with fungal isolation in the lymph nodes, spleen and liver at the 30 and 60 days postinoculation. No significant differences were observed between cytopathology, histopathology and fungal culture during the different phases of the disease. Therefore, cytological examination was considered a simple, rapid and inexpensive diagnostic method at all stages of this disease. Topic 4: PARASITIC DISEASES P-21 Comparison of a permethrin-pyriproxyfen spray and fipronil spot-on used alone in a therapeutic trial for the diagnosis of canine flea allergy dermatitis P. JASMIN, M. BRIGGS, H. SCHROEDER and A. SANQUER Virbac SA, Carros, France; Newlands Veterinary Clinic, Cape Town, South Africa; Willow Park Veterinary Hospital, Lynnwood Ridge, South Africa This randomized clinical trial compared two parasiticides [permethrin-pyriproxyfen spray (Duogard Ò Spray/ Duowin Ò Spray) and fipronil spot-on (Frontline Ò Spoton)] for use in the diagnosis of canine flea allergy dermatitis (FAD). Dogs with a flea infestation, minimum FAD clinical score and a positive intradermal test were treated with either one of the parasiticides applied three times at 1-week intervals. It was required that the dogs live partially outdoors, with at least one nontreated in-contact pet, and an environment that had not been treated within the previous year. Both groups were compared for clinical improvement and the number of dogs showing a positive response to the therapeutic test (defined as > 75% clinical improvement). Twenty-seven dogs were included; two were lost to follow-up, 12 were treated with the spray and 13 with the spot-on. Mean scores for pruritus and lesions decreased rapidly in the permethrin-pyriproxyfen spray group, but more slowly in the fipronil spot-on group. There was a significant difference between the two groups at all consultation time points (P < 0.05). After 3 weeks, responses were positive in all dogs treated with the permethrin-pyriproxyfen spray, and significantly higher than with the fipronil spot-on (P < 0.05). The sensitivity of the therapeutic trial was 100% for the spray and 31% for the spot-on. Negative predictive values varied from 14 to 65% with the spot-on and were 100% with the spray. No adverse effects were observed. The permethrin-pyriproxyfen spray can be recommended as a useful product to perform a therapeutic trial in confirming a diagnosis of FAD in dogs. Funding: Virbac SA. P-22 Mites of interest in canine and feline dermatology: evaluation of their survival and structural integrity in oil preparations for microscopy G. A. MANIGOT Dermlink Buenos Aires, Buenos Aires, Argentina Diagnostic dilemmas often arise when veterinary practitioners suspect a parasitic skin disease but direct microscopy of collected samples are negative. It has been suggested that the delay between collection of the sample in the veterinary practice and evaluation at the referral laboratory results in parasite lysis. This study was performed to determine if delay in processing the samples may lead to negative results. A further aim was to determine if using other oil media (when mineral oil was not available) would result in parasite modification. Onehundred samples of skin scrapings and otic smears in several oil media from dogs and cats diagnosed as positive for several mites of dermatological importance were kept under room conditions and evaluated over a 21-day period for mite survival and retention of recognizable structural features. Mineral oil, kitchen grade olive oil, pharmaceutical grade baby oil and glycerin were used as oily media to process the samples. At the end of the study, mite survival rate was established between 2 and 5 days. Sarcoptes and Otodectes mites survived the longest. After 21 days of observation, all parasites and their eggs still remained recognizable, no matter which oil medium was used. It was concluded that a delay in the submission of samples does not result in negative findings.

48 P-23 Clinical and epidemiological characteristics of 153 cases of Sarcoptic acariosis in dogs P. BOURDEAU, L. ARMANDO and A. MARCHAND National Veterinary School of Nantes, Nantes, France Although considered a classical skin disease, Sarcoptes scabiei infestation is uncommon in canine dermatology in some countries. Information concerning the frequency, epidemiology and clinical signs is scarce. The aim of this study was to analyse information obtained from our dermatology clinic from three successive periods: (A) retrospectively (1983 1992), (B) prospectively (1993 1996), and (C) prospectively (1993 2002). From a total of 12,112 dogs, the number of proven cases was only 153 (1.26%, range of 0.8 1.45%, depending on the study). Average age was 2.3 2.7 years (range 3 weeks 16 years) with 52.4 61.6% of dogs being <1-year old. Pruritus was severe in 61 77% of cases. Otopedal reflex was positive in 43 56% (positive in 4% of the general population in B) and was significantly more common when skin lesions were extensive. Contagion to other dogs was seen in 5 33% of cases with human involvement in 4 20% of cases. No sex predisposition was noted but breed predilection may exist. Lesions were noted on the ventrum (81%), ear pinnae (71.8%), elbows (66%) and distal legs (45%). Although the diagnosis was confirmed by different methods, Sarcoptes mites were isolated and identified in 22.8 50% of cases. Fluctuations in incidence of cases may occur from year to year, but no change in the overall frequency was observed during this 20-year period in the area served by our clinic. P-24 Efficacy of selamectin in canine scabies and ear mite infestation in Korea T. H. OH, W. S. OH, S. R. KIM, H. Y. OH, W. P. HOH, A. Y. JEONG, H. H. JEONG and S. J. PARK College of Veterinary Medicine, Kyungpook National University, Daegu, Korea; College of Veterinary Medicine, Chungnam National University, Daejeon, Korea A randomized and controlled field study was performed in canine patients to evaluate the efficacy of selamectin in the treatment of naturally occurring Sarcoptes scabiei and Otodectes cynotis infestations in dogs. A total of 227 dogs from six veterinary practices in South Korea were included. Dogs were randomly assigned to treatment with selamectin or a positivecontrol product. Selamectin was administered as a unit dose providing a minimum of 6 mg/kg in a topical preparation applied to the skin in a single spot on days 0 and 30 [S. scabiei (n =113) and O. cynotis (n =114)]. The presence of parasites was assessed before treatment and at 14, 30 and 60 days after the initiation of treatment. The animals were evaluated clinically at each assessment period. Based on skin scrapings, the efficacy of selamectin against S. scabiei infestations on dogs was >95% by day 30, and 100% by day 60. Against O. cynotis, selamectin eliminated mites in 100% of dogs by day 60. However, clinical signs of pruritus, erythema, scale, and crusted papules did not diminish concomittantly with resolution of S. scabiei in skin scrapings. The positive-control products achieved similar results. Therefore, selamectin was safe and effective against sarcoptic mange and ear mites in dogs. Funding: Pfizer Animal Health. P-25 Sensitivity of deep skin scrapings, hair pluckings and exudate microscopy in the diagnosis of canine demodicosis M. N. SARIDOMICHELAKIS, A. F. KOUTINAS, R. FARMAKI and L. LEONTIDES School of Veterinary Medicine, Aristotles University of Thessaloniki, Greece; School of Veterinary Medicine, University of Thessaly, Karditsa, Greece Diagnosis of canine demodicosis is mainly based on mite detection in deep skin scrapings (DSS). Hair pluckings (HP) are also helpful, whereas the diagnostic value of exudate microscopy (EM) has never been assessed. The objective of this study was to compare the diagnostic sensitivity of DSS, HP and EM in the various clinical forms of canine demodicosis. A total of 68 dogs with localized (45.6%) or generalized (54.4%) demodicosis, juvenile-onset (92.6%) or adult-onset (7.4%) demodicosis were included. Pyoderma was present on 41.2% of cases and absent in 58.8% of cases. One lesion per dog was selected for obtaining one HP, DSS and EM sample, the latter only in 14 cases demonstrating visible exudate. Mineral oil DSS and EM microscopy was focused on a 2.2 2.2 mm area, while a total of 100 hair shafts were evaluated by trichogram. At least one parasite was seen in 83.8% of HP, 98.5% of DSS and 100% of EM slides. The sensitivity of HP was higher in the generalized (P =0.001) and pyoderma-affected (P =0.021) cases compared to the localized and noninfected cases. A higher sensitivity of DSS compared to HP was demonstrated in the localized (P =0.004) and noninfected (P = 0.002) cases. In conclusion, HP should be viewed as a useful diagnostic tool in generalized and infected canine demodicosis, while EM is useful in all deep pyoderma-associated cases.

49 P-26 New aspects of canine demodicosis and the possibility of treatment with selamectin N. TARPATAKI, E. KADOCSA and K. VÖRO S Faculty of Veterinary Sciences, Szent Istva n University, Budapest, Hungary The purpose of this study was to evaluate 55 clinical cases of canine demodicosis and to compare the results of treatment using amitraz (solution), selamectin (spot-on), ivermectin (injection) and cythioate (oral tablets). Data from the 55 cases was collected and evaluated after clinical and microbiological examination. Treatment was selected depending on the severity of demodicosis and compliance of the owner. The cases were followed for 12 months and the status of the patients was grouped on two levels: recovered (58%), or relapsed (42%). Five dogs (9%) were euthanized. The disease was commonly diagnosed in purebred dogs. Demodicosis was more common in dogs under 2 years of age (65%), in males (64%), and in the short-haired breeds (75%). Demodicosis was generalized in 73% of cases, localised in 23% and affected the feet (pododemodicosis) in 4% of cases. Recovery was the highest in dogs between 1 and 2 years of age (73%), and in the localized cases (92%). Nonspecific treatment with glucocorticoids prior to the diagnosis lowered the rate of recovery (4%), but treatment with glucocorticoids for proven atopic dermatitis improved the rate of recovery (41%). All drugs (amitraz, selamectin, cythioate) administered for the localized form were effective (100% recovered). Recovery in generalized demodicosis was 60% using ivermectin, 55% using amitraz, 44% with the combination of amitraz and selamectin (two treatments with amitraz followed by selamectin), and 43% in cases where selamectin was used alone. Funding: Pfizer Animal Health. P-27 Characteristics of Dermacentor reticulatus and Ixodes ricinus infestations in 191 dogs in France P. BOURDEAU, T. XIMENES and O. BEZIADE National Veterinary School of Nantes, Nantes, France; Clinique Ve te rinaire, Gue rande, France; Clinique Ve te rinaire, Saint-Gaudens, France The aim of this study was to evaluate the distribution of ticks on the body and characteristics of tick populations and their consequences. The studies were conducted in France in two clinics (A and B) over 10 months. Dogs were carefully examined for the presence of ticks (all dogs in group A, and 150 dogs per month in group B). Information on hair coat (long, medium, short), pruritus, number of ticks, distribution of ticks on the body, and identification of ticks was collected. An intradermal test (Dermatophagoides farinae, D. pteronyssinus) was performed on most of the tick-positive dogs in group A. In group A, 87 dogs (4.5%) were infested with ticks, while in group B, 104 dogs (9%) were infested. There was no significant difference in infested dogs compared with control dogs with regard to coat length. The level of infestation was generally very low: <5 ticks in 81.5%, 5 9 ticks in 12.2%, and 10 in 6.3% of infested dogs. Of 528 ticks collected, Dermacentor reticulatus was the most abundant (67%), followed by Ixodes ricinus (32.9%) and Rhipicephalus sanguineus (0.1%). Ticks were found mainly on the head (41%) and neck (52.9%), and less frequently on the trunk, ventrum, legs, perianal and tail areas. Twenty-eight per cent of infested dogs were pruritic compared to only 10% in ectoparasite-free dogs. In 65 intradermal tests, 23% were positive to D. farinae and 6% to D. pteronyssinus. P-28 Efficacy of selamectin spot-on formulation against Neotrombicula autumnalis in eight cats F. LEONE and F. ALBANESE Clinica Veterinaria Adriatica, Senigallia (AN), Italy; Clinica Veterinaria L Arca, Napoli, Italy The purpose of this study was to evaluate the efficacy of a topical formulation of selamectin in the treatment of naturally acquired Neotrombicula autumnalis infestations in eight indoor/outdoor cats. Selamectin was administered topically once at a dosage of 6 mg/kg. Clinical examination and visualization of mites by microscopic examination of skin scrapings were performed after 2, 30 and 60 days. By day 2, clinical signs had subsided and all mites were dead on microscopic examination. No signs of recurrence were apparent at follow-up. Under the conditions of our study, topical selamectin was safe and 100% effective against natural infestations of Neotrombicula autumnalis. To our knowledge, this is the first report demonstrating the efficacy of selamectin against Neotrombicula autumnalis. P-29 Pustular dermatosis caused by fire ant (Solenopsis invicta) stings in a dog L. G. CONCEICAO, V. HADDAD, F. H. LOURES, M. E. LEAL and F. CLARE Universidade Federal de Vicosa, Vicosa, Minas Gerais, Brazil; Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, Sao Paulo, Brazil Solenopsis invicta (lavape s ants or imported fire ants) are insects that belong to the Hymenoptera order and Formicoidea superfamily. Native to Brazil, this aggressive ant species is today disseminated throughout the Americas. In humans, the sting of the fire ant causes an urticarial papule that progresses within 24 h to a highly pruritic, sterile pustular dermatitis. Children and alcoholics are more often affected, presenting with massive injuries. Subcorneal neutrophilic pustular dermatitis is the usual histopathological finding. One study has reported that fire ant stings in dogs leads to a vertically oriented necrotizing dermatitis with collagen degeneration. This abstract reports a natural fire ant-induced dermatosis in a dog. We report a female, intact, young-adult, mixed-breed dog presented for routine check-up. Physical examination revealed several grouped, nonfollicular, circumscribed abdominal and inguinal pustules. The pustules were relatively tense, circumscribed by a slight erythematous halo, and several dead fire ants were attached to the skin and hairs. No bacteria were seen on cytological examination. Histopathological examination revealed an intraepidermal neutrophilic pustule and dense dermal interstitial, predominantly neutrophilic, infiltration, which extended to the panniculus. Collagen degeneration and edema were also observed. This case documents that fire ant stings can produce pustular intraepidermal dermatitis in dogs similar to that observed in humans.

50 P-30 Use of fipronil for treatment of Lynxacarus radovskyi in outdoor cats in Rio de Janeiro (Brazil) F. CLARE, R. M. L. C. MELLO, T. V. BASTOS, C. LESSA and L. G. CONCEIC A O Curso de Medicina Veterina ria, Universidade Castelo Branco, Rio de Janeiro, Brazil The aim of this study was to report 20 cases of Lynxacarus radovskyi infestation in 20 cats. Lynxacarus radovskyi was diagnosed by physical examination and microscopic examination of plucked hair where countless adult mites and immature stages were identified. Microscopic examination was repeated every 7 days. The animals were observed daily for the first 7 days, and again on day 15 and day 30 for mites, pruritus and clinical signs. Clinical signs included salt-andpepper appearance to a dull and opaque hair coat, alopecia of the dorsal and lateral areas of the hind limbs, and evidence of self-mutilation. One case had papules and crusts of the hind limbs. The pruritus was mild in animals with low infestation, but very intense in those with a heavy infestation. Most mites occurred in the lateral area of the hind limbs and sacral area. Twenty of 100 animals had confirmed L. radovskyi infestation. Therefore, we suggest that the transmission occurs through fomites. Treatment consisted of 0.5 ml application of Fipronil Ò (Frontline TopSpot) per cat, with resolution in 100% of cases by day 15. Funding: Merial. P-31 Anal sacs myiasis: a case report S. MOHITMAFI Faculty of Veterinary Medicine, Islamic Azad University, Karaj, Tehran, Iran A 3-year-old male Doberman dog was presented with signs of anorexia, constipation and dyschezia. Clinical examination revealed abdominal pain, dehydration, swelling around the anus, and a small wound on the left side of the perineal region where myiasis was clearly observed. Haematological tests showed increased packed cell volume, and an increased white cell count with a mild neutrophilia and eosinophilia. Radiography showed severe constipation and obstruction of the rectum due to external compression from a soft tissue swelling. Cannulation of the anal sacs produced a small amount of sanguinous fluid. With a diagnosis of severe impaction in the anal sacs, surgery was performed and a large number of larvae were observed inside the anal sacs. Anal sacs myiasis was diagnosed. No connections were found between the anus and the rectum at surgery. Larvae and the necrotized tissue were all surgically removed and after 8 days of supportive treatment, the patient was released from hospital. Topic 5: VIRAL DISEASES P-32 Giant cell dermatosis in a FeLV-infected cat: detection of the virus in infected skin by immunohistology and PCR C. FAVROT, P. GREST, M. MELI, A. KIPAR, S. WILHELM and N. ZAUGG Faculty of Veterinary Medicine, University of Zurich, Zurich, Switzerland; Faculty of Veterinary Science, University of Liverpool, Liverpool, UK A 3-year-old castrated male domestic short-haired cat was presented with a dermatosis of 3-months duration. Physical examination revealed well demarcated ulcerative lesions that affected the head, limbs and paws. The cat was also depressed, febrile and anaemic. Previous history included vaccination for FeLV twice during the first year of life, and again in the second year. A staphylococcal infection was diagnosed by cytological and bacteriological examination. An ELISA for FeLV antigen was positive. Additionally, histological examination of the skin revealed an ulcerative dermatitis with folliculitis, dyskeratotic keratinocytes and syncytium formation within the epidermis and sebaceous glands. A tentative diagnosis of FeLV-associated giant cell dermatosis with secondary staphylococcal folliculitis was made and the cat was started on cephalexin. Despite the treatment and a marked, but temporary, improvement of the skin lesions, the general condition deteriorated and the cat was euthanized. Necropsy was not performed. Immunohistological staining with monoclonal antibodies against the envelope protein gp70 and the group-specific protein p27 of FeLV revealed positive cells in the epidermis and the sebaceous glands. Additionally, PCR analysis was performed on lesional skin. DNA was isolated from deparaffinized tissues and a 131 bp long proviral DNA fragment was amplified by realtime PCR. Giant cell dermatosis is a rare consequence of FeLV infection in cats. Cases have already been described and confirmed by immunohistology. In man, retroviral infections are also known to induce syncytium formation.