Veterinary Dermatology 2003, 14, 153157

Comparative efficacies of oral ketoconazole and terbinafine for

reducing Malassezia population sizes on the skin of Basset Hounds

Blackwell Publishing Ltd.

JACQUES GUILLOT*, EMMANUEL BENSIGNOR, FRANOIS JANKOWSKI,

WOLFGANG SEEWALD, REN CHER METTE* and JEAN STEFFAN
*Equipe de Mycologie, UMR Biologie Molculaire et Immunologie Parasitaires et Fongiques,
Ecole Nationale Vtrinaire dAlfort, Maisons-Alfort, France
Dermatology Referral Service, Veterinary Clinic, Paris, France
Novartis Sant Animale, Rueil Malmaison, France
Animal Health, Basel, Switzerland
(Received 16 April 2002; accepted 31 January 2003)

Abstract 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 kg1, 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 kg1,
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.
Keywords: Basset Hounds, dog, ketoconazole, Malassezia, terbinafine.

INTRODUCTION
The lipophilic yeast Malassezia pachydermatis is now
recognized as a normal inhabitant of canine skin and
mucosae.13 However, M. pachydermatis may become a
pathogen when alteration in the skin surface microclimate
or host defence occurs. Otitis externa, as well as seborrhoeic dermatitis, in dogs are frequently associated with
cutaneous proliferation of M. pachydermatis.1,4 Predisposing factors include allergic diseases, cornification
disorders, bacterial skin infections as well as long-term
glucocorticoid or antibiotic administration.1,4,5 Several systemic and topical regimens have been proposed
for the treatment of canine dermatitis associated with
abnormally high population sizes of M. pachydermatis.
Oral ketoconazole at 10 mg kg1 once daily (or 5 mg kg1
twice daily) is the most widely used and least expensive of orally administered azole drugs. However, this
drug is not licensed for use in dogs world-wide. In
addition, adverse effects, such as anorexia and raised
liver enzymes have been reported.6 Terbinafine is a

new antifungal drug that inhibits ergosterol biosynthesis

by interfering with the enzyme squalene epoxidase.7
It has shown excellent efficacy against dermatophyte
infections, especially onychomycoses in humans.6
Similar results were obtained in natural810 or experimental11,12 dermatophytoses in animals. A recent investigation of in vitro activity of four antifungal drugs
against Malassezia species demonstrated that M. pachydermatis was highly susceptible to terbinafine.13
The purpose of the study reported here was to compare the efficacy of oral ketoconazole and terbinafine
for reducing population sizes of Malassezia yeasts on
the skin of healthy Basset Hounds. The dogs all
belonged to the same kennel in south-west France and
had been previously used in a study whose purpose was
the comparison of two sampling techniques to assess
quantity and distribution of the Malassezia yeasts on
canine skin.14

MATERIALS AND METHODS

This study was a monocentric, randomized, blinded and
controlled clinical field trial. The study design details
are described below.

Conflict of interest: this study was sponsored by Novartis Animal

Health.
Correspondence: J. Guillot, ParasitologieMycologie, Ecole Nationale
Vtrinaire dAlfort, 7, avenue du Gnral de Gaulle, 94704 MaisonsAlfort Cedex, France. E-mail: jguillot@vet-alfort.fr
2003 European Society of Veterinary Dermatology

Animals
Twenty-one Basset Hounds, aged between 1 and 9
years old, were included in the trial. Three were male
153

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J. Guillot et al.

and 18 were female. The animals were all privately

owned dogs from the same kennel. All dogs were in
good general health with no history of pruritus or
skin disease and had no inflammatory skin disease,
except a mild greasy seborrhoea in eight animals
which did not require treatment. None of the dogs had
been treated with a systemic or topical antifungal drug
in the previous 30 days before the beginning of the
experiment.
Therapeutic protocol
Dogs were divided into three subgroups according to
Malassezia population sizes assessed in four cutaneous
sites (left pinna, left axilla, umbilical region and neck)
and presence or absence of seborrhoea 2 weeks before
the beginning of the trial (day 14). Each subgroup of
seven dogs consisted of animals with comparable
Malassezia population sizes on the skin. Individual
dogs within each subgroup were then randomized to
three therapeutic groups (randomised complete group
design). Sex and age were not taken into account for
randomization. The dogs were not separated during
the experiment. The seven Basset Hounds in the first
group were treated by oral administration of ketoconazole (Ketofungol 200 mg, Janssen-Cilag) at
10 mg kg1, every 24 h with food, for 3 weeks. The
seven dogs in the second group were treated by oral
administration of terbinafine (Lamisil 250 mg,
Novartis) at 30 mg kg1, every 24 h with food, for
3 weeks. The exact dosage of ketoconazole or terbinafine to be administered was calculated for each dog on
the basis of individual body weight. No other topical
or systemic therapy was allowed during the period of
the experiment. The seven remaining dogs were not
treated and considered as controls.
Determination of Malassezia counts at four anatomic
sites
At days 14, 0, 7, 14 and 21, samples were collected
from four different anatomic areas (left pinna, left
axilla, umbilical region and neck) with contact plates
as already described.5,14 One count was carried out at
each site in each dog. Contact plates were then incubated at 32 C for 4 days before counting. Malassezia
yeasts or any other fungal species were recognized by
microscopic examination of a wet mount from 10
representative colonies and by physiological tests
including catalase, -glucosidase and Tween assimilation
tests.15 Malassezia colonies were counted until 1000 by
an investigator who was not aware of the site of sampling or the treatment group. If more than 1000 colonies were present on the plate, the investigator did not
count further.
Statistics
The randomization of the dogs in the treatment groups
was generated using the PLAN procedure of software (Version 8, SAS Institute Inc., Cary, NC, USA).
Yeast population sizes were compared between the
three groups using a repeated measurements analysis-

of-variance model, allowing for differences between

D7, D14 and D21, and also between the four sites.
Baseline values were included in the model as a covariate. Using linear contrasts, the three treatment groups
were then compared; P-values were adjusted for multiple testing using the TukeyKramer correction. A
P-value < 0.05 was considered significant. Additional
analyses were performed for the data from one site
only, or from one day only.
Yeast population counts (c) were reported only up
to a maximum number of 1000; higher values were
reported as > 1000. Because these counts were not
normally distributed, transformation had to be applied
before subjecting the data to the analysis of variance.
The following transformation (lc) yielded a good
approximation to the normal distribution:
lc = log10(c + a), when c < 1000;
lc = log10(b), when c > 1000.
When > 1000 colonies were found, c was standardized
to 1000. Several values for a and b were tested in order
to obtain an approximate normality of the residuals of
the model. In the final analysis a value of 20 was used
for a and 1200 for b. After transformation the data set
was normally distributed.

RESULTS
From day 0 to day 21, Malassezia yeasts were isolated
from the four anatomic sites in all the dogs. At the
beginning of the study, yeast counts exceeded 1000
colonies per plate in about one third of the samples.
All Malassezia colonies were initially isolated on a
lipid-supplemented medium but subcultures were
systematically obtained on Sabouraud dextrose agar
without any lipid supplementation. The species
M. pachydermatis was thus identified in all dogs and at
all anatomic sites. No lipid-dependent Malassezia
yeasts, or filamentous fungi could be isolated.
Initial Malassezia counts were comparable in the
three groups. There were, respectively, three, two and
three dogs with seborrhoea in the control, ketoconazole and terbinafine groups, respectively. Yeast counts
in ketoconazole and terbinafine treated dogs and in
controls are shown in Fig. 1. Statistical tests were performed with the full data set, but also with data sets
corresponding to a single anatomic site (left pinna, left
axilla, umbilical region and neck), or to a single sampling date (7, 14 or 21). The estimated difference of
both therapeutic groups (with the transformed scale)
to the control group is shown in Table 1. P-values for
pairwise comparisons are shown in Table 2. The results
clearly confirmed the efficacy of oral ketoconazole for
reducing Malassezia population sizes on canine skin.
The overall difference to the control group on a logarithmic scale was 1.00, which corresponded to a
decrease in yeasts count of a factor of about 2.70.
Highly significant differences were detected with the

2003 European Society of Veterinary Dermatology, Veterinary Dermatology, 14, 153157

Efficacy of oral terbinafine for reducing Malassezia population sizes

155

control group from day 7 to day 21 and for each sampling site (Table 1). Terbinafine was also proven to have
in vivo antifungal activity against Malassezia yeasts.
The overall difference from the control group on a logarithmic scale was 0.51, which corresponded to a
decrease in yeasts count of a factor of 1.6. Significant
differences with the control group were not detected
site per site but only in total (Table 1). There was no
significant difference between ketoconazole and terbinafine (P = 0.0530). Both treated groups showed a
statistically lower number of yeasts on days 14 and 21
compared with the control, but not on day 7 (Table 2).
No adverse effect was observed in either ketoconazoleor terbinafine-treated dogs.

DISCUSSION

Figure 1. Yeast counts in controls (C), ketoconazole-treated (K) and

terbinafine-treated (T) dogs from day 0 to day 21. Data from the four
sampling sites are combined. Boxes range from lower to upper
quartile, with a circle at the mean; vertical lines extend to the
minimum and maximum value, with outliers shown as asterisks.

Previous investigations clearly demonstrated that the

skin of Basset Hounds is extensively colonized by
Malassezia yeasts.14,16 In fact, both the frequencies of
isolation and populations sizes of the yeasts were
found to be significantly higher at skin and mucosal
sites of Basset Hounds when compared with those of
healthy dogs of various other breeds.16 In Basset
Hounds, yeast proliferation may be allowed by a primary keratinization defect leading to erythema and
greasy exudation.17 The presence of deep skin folds in
Basset Hounds may also account for high Malassezia
populations. Regardless of the mechanisms which
allow the proliferation of the yeasts, Basset Hounds
represent good models for the evaluation of in vivo
anti-Malassezia activity of topical or systemic drugs.
Bond et al.2 selected 33 Basset Hounds with Malasseziaassociated seborrhoeic dermatitis to compare the
clinical and antimicrobial activities of two topical
treatments.18 They demonstrated that a superior clinical performance was related to low post-treatment
microbial counts in a miconazolechlorhexidine
group, whereas higher populations were present in

Table 1. Estimated differences in the mean yeast count between therapeutic groups and the control group. Sampling sites were: left auricular
pinna (A), neck (B), left axilla (C), and umbilical region (D). Transformed data were used (transformed count = log 10(count + 20), when yeast
count was < 1000, and transformed count = log10 (1200), when yeast count was > 1000)
Therapeutical group

All data*

Site A

Site B

Site C

Site D

Day 7

Day 14

Day 21

Terbinafine
Ketoconazole

0.52
1.00

0.76
1.49

0.53
0.92

0.52
0.82

0.41
0.82

0.35
0.60

0.80
1.24

0.63
1.14

*Pooled data from all sites and all days (7, 14 and 21). Pooled data on all days. Pooled data on all sites.

Table 2 P-values for pairwise comparisons in yeast count between therapeutic and control groups. Sampling sites were: left auricular pinna (A),
neck (B), left axilla (C), and umbilical region (D). In each column, adjustment for multiple testing was done using the TukeyKramer correction.
P-values < 0.05 are considered significant and are indicated in bold

Control vs. ketoconazole

Control vs. terbinafine
Ketoconazole vs. terbinafine

All Data*

Site A

Site B

Site C

Site D

Day 7

Day 14

Day 21

0.0016
0.1041
0.1405

0.0072
0.1991
0.2318

0.0313
0.2701
0.4791

0.0265
0.1900
0.5695

0.1084
0.5491
0.5371

0.1428
0.4874
0.6943

0.0012
0.0323
0.3033

0.0018
0.0849
0.1870

*Pooled data from all sites and all days (7, 14 and 21). Pooled data on all days. Pooled data on all sites.
2003 European Society of Veterinary Dermatology, Veterinary Dermatology, 14, 153157

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J. Guillot et al.

selenium sulfide-treated dogs. This study was the first

to prove that yeast population sizes should be considered as of major significance in the development of
seborrhoeic dermatitis in dogs. In the present
investigation, the selected dogs had no dermatitis and
we could not compare antifungal activity to clinical
improvement. Ketoconazole showed the higher efficacy with a significant reduction of yeast populations.
Oral terbinafine was also proven to have anti-Malassezia
activity. The reduction factor in terbinafine-treated
dogs was less than that obtained in ketoconazoletreated dogs (1.6 vs. 2.7) but the difference was not
statistically significant. As a consequence, oral terbinafine at 30 mg kg1 every 24 h should be evaluated as a
possible alternative to azole derivatives for the management of Malassezia-associated dermatitis in dogs.
The main advantage of terbinafine is represented by a
high safety margin in man.7,19 Because terbinafine has
no inhibitory effect on cytochrome P450 systems, it is
more selective than azole derivatives such as ketoconazole. In rodents and dogs, no embryonic or foetal toxicity or teratogenicity has been reported. Furthermore,
terbinafine has a relatively low potential for drug interactions.7,20 Oral terbinafine is currently used with success for the treatment of onychomycosis in humans.19
Terbinafine continues to have a clinical effect after cessation of treatment, due to residual tissue levels of the
drug.7 Cases of dermatophytosis in dogs, cats and
rodents have also been treated with terbinafine without
occurrence of adverse effects.810 Faergemann recently
suggested that oral terbinafine should be evaluated for
the treatment of Malassezia dermatitis in humans.21 In
vitro susceptibility tests demonstrated the efficacy of
terbinafine against all Malassezia species.13,22 However, strains of M. furfur, M. globosa and M. obtusa
were more resistant to terbinafine than the other
Malassezia species including M. pachydermatis, which
is the only species present on canine skin. The yeast
M. sympodialis that may be isolated from the skin and
mucosae in cats was proven to be highly susceptible to
terbinafine. Further clinical investigations are needed
for the definitive demonstration of the usefulness of
terbinafine in nondermatophyte cutaneous infections
such as Malassezia-associated dermatitis.
REFERENCES
1. Scott, D.W., Miller, W.H., Griffin, C.E. Fungal skin diseases. In: Muller and Kirks Small Animal Dermatology,
6th edn. W.B. Saunders, Philadelphia, 2000: 363 74.
2. Bond, R., Saijonmaa-Koulumies, L.E.M., Lloyd, D.H.
Population sizes and frequency of Malassezia pachydermatis
at skin and mucosal sites on healthy dogs. Journal of
Small Animal Practice 1995; 36: 14750.
3. Guillot, J., Bond, R. Malassezia pachydermatis: a review.
Medical Mycology 1999; 37: 295 306.
4. Plant, J.D., Rosenkrantz, W.S., Griffin, C.E. Factors
associated with a prevalence of high Malassezia pachydermatis numbers on dog skin. Journal of the American
Veterinary Medical Association 1992; 201: 879 85.

5. Bond, R., Collin, N.S., Lloyd, D.H. Use of contact plates

for the quantitative culture of Malassezia pachydermatis
from canine skin. Journal of Small Animal Practice 1994;
35: 6872.
6. Hill, P.B., Moriello, K.A., Shaw, S.E. A review of systemic antifungal agents. Veterinary Dermatology 1995; 6:
5966.
7. Gupta, A.K., Sauder, D.N., Shear, N.H. Antifungal
agents: an overview: part II. Journal of the American
Academy of Dermatology 1994; 30: 91133.
8. Castanon-Olivares, L.R., Manzano-Gayosso, P.,
Lopez-Martinez, R. et al. Effectiveness of terbinafine in
the eradication of Microsporum canis from laboratory
cats. Mycoses 2001; 44: 957.
9. Chen, C. The use of terbinafine for the treatment of dermatophytosis. Veterinary Dermatology 2000; 11 (Suppl.
1): 41.
10. Mancianti, F., Pedonese, F., Millanta, F. et al. Efficacy of
oral terbinafine in feline dermatophytosis due to Microsporum canis. Journal of Feline Medicine and Surgery
1999; 1: 3741.
11. Mieth, H., Leitner, I., Meingassner, J.G. The efficacy of
orally applied terbinafine, itraconazole and fluconazole
in models of experimental trichophytoses. Journal of
Medical and Veterinary Mycology 1994; 32: 1818.
12. Petranyi, G., Meingassner, J.G., Mieth, H. Activity of
terbinafine in experimental fungal infections of laboratory animals. Antimicrobial Agents and Chemotherapy
1987; 31: 155861.
13. Gupta, A.K., Kohli, Y., Li, A. et al. In vitro susceptibility
of the seven Malassezia species to ketoconazole, voriconazole, itraconazole and terbinafine. British Journal of
Dermatology 2000; 142: 75865.
14. Bensignor, E., Jankowski, F., Seewald, W. et al. Comparison
of two sampling techniques to assess quantity and distribution of Malassezia yeasts on the skin of healthy Basset
Hounds. Veterinary Dermatology 2002; 13: 23741.
15. Guillot, J., Guho, E., Lesourd, M. et al. Identification
of Malassezia yeasts: a practical approach. Journal de
Mycologie Mdicale 1996; 6: 10310.
16. Bond, R., Lloyd, D.H. Skin and mucosal populations of
Malassezia pachydermatis in healthy and seborrhoeic
Basset Hounds. Veterinary Dermatology 1997; 8: 1016.
17. Power, H.T., Ihrke, P.J., Stannard, A.A. et al. Use of
etretinate for treatment of primary keratinization disorders (idiopathic seborrhoea) in Cocker Spaniels, West
Highland White Terriers and Basset Hounds. Journal of
the American Veterinary Medical Association 1992; 201:
41929.
18. Bond, R., Rose, J.F., Ellis, J.W. et al. Comparison of two
shampoos for treatment of Malassezia pachydermatisassociated seborrhoeic dermatitis in Basset Hounds.
Journal of Small Animal Practice 1995; 36: 99104.
19. McClellan, K.J., Wiseman, L.R., Markham, A. et al. An
update of its use in superficial mycoses. Drugs 1999; 58:
179202.
20. Shear, N., Drake, L., Gupta, A.K. et al. The implications
and management of drug interactions with itraconazole,
fluconazole and terbinafine. Dermatology 2000; 201:
196203.
21. Faergemann, J. Treatment of seborrhoeic dermatitis with
oral terbinafine? the Lancet 2001; 358: 170.
22. Petranyi, G., Meingassner, J.G., Mieth, H. Antifungal
activity of the allylamine derivative terbinafine in vitro.
Antimicrobial Agents and Chemotherapy 1987; 31: 13658.

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Rsum Le but de cette tude tait de comparer lefficacit du ktoconazole et de la terbinafine pour diminuer
les populations de Malassezia sur la peau du chien. Ving et un Basset Hound ont t distribus au hasard dans
trois groupes de 7 chiens, en fonction des populations de Malassezia. Les chiens du premier groupe ont t traits
par ladministration orale de ktoconazole (Ketofungol 200 mg, Janssen-Cilag) 10 mg/kg, une fois par jour
au moment du repas, pendant trois semaines. Les chiens du deuxime groupe ont reu de la terbinafine (Lamisil
250 mg, Novartis), 30 mg/kg, une fois par jour avec le repas pendant 3 semaines. Les chiens du dernier groupe
ont servi de contrle. Les populations de Malassezia ont t values par lutilisation de boites de contact sur 4
sites cutans, J7, J14 et J21. Le ktoconazole et la terbinafine ont t efficaces pour diminuer les populations
de Malassezia, sans diffrence significative entre les deux molcules. Des tudes supplmentaires sont indiques
pour dterminer si lutilisation de la terbinafine par voie orale est intressante pour traiter les cas de dermatite
Malassezia chez le chien.
Resumen El objetivo del actual estudio era comparar la eficacia del ketoconazol y de la terbinafina orales para
reducir el tamao de las poblaciones de las levaduras de Malassezia en piel canina. Veintin Basset Hounds
fueron seleccionados al azar en tres grupos de 7 segn las poblaciones de Malassezia. Los perros en el primer
grupo fueron tratados mediante la administracin oral de ketoconazol (Ketofungol 200 mg, Janssen-Cilag) a
dosis de 10 mg kg1, cada 24h con la comida, durante tres semanas. Los perros del segundo grupo fueron tratados
mediante la administracin oral de la terbinafina (Lamisil 250 mg, Novartis) a dosis de 30 mg kg1, cada 24h
con la comida, durante tres semanas. Los siete perros restantes fueron utilizados como controles. Los tamaos
de las poblaciones de Malassezia fueron determinados por medio de la utilizacin de placas de contacto en 4
sitios cutneos, en los das 7, 14 y 21. Tanto el ketoconazol como la terbinafina fueron eficaces en la reduccin
de los niveles basales de los organismos de Malassezia sin diferencia significativa entre las dos drogas. En futuros
estudios, la terbinafina oral debera evaluarse para el control de casos caninos de dermatitis por Malassezia.
Zusammenfassung Das Ziel dieser Studie war, die Wirksamkeit von oralem Ketoconazol und Terbinafin in der
Reduktion von Malassezia Hefen auf Hundehaut zu vergleichen. Einundzwanzig Bassets wurden nach ihrer
Malassezia Population randomisiert in drei Gruppen mit 7 Hunden aufgeteilt. Hunde in der ersten Gruppe
erhielten fr drei Wochen 10 mg kg1 Ketoconazol (Ketofungol 200 mg, Janssen-Cilag) oral alle 24 h mit Futter.
Hunde der zweiten Gruppe wurden drei Wochen lang mit 30 mg kg1 Terbinafin (Lamisil 250 mg, Novartis)
oral mit Futter alle 24 h behandelt. Die restlichen sieben Hunde dienten als Kontrolle. Malassezia Populationsdichte wurde mit Kontaktplatten nach sieben, 14 und 21 Tagen an vier Hautstellen bewertet. Sowohl Ketoconazol als auch Terbinafin reduzierten das Basisniveau der Malassezia Organismen, zwischen den zwei
Medikamenten bestand kein signifikanter Unterschied. Orales Terbinafin sollte in weiteren Studien fr die
Behandlung der Malassezia-Dermatitis des Hundes bewertet werden.

2003 European Society of Veterinary Dermatology, Veterinary Dermatology, 14, 153157