Veterinary Research Communications, 20 (1996) 161-166

Copyright 6 Kluwer Academic Publishers bv - Printed in the Netherlands

ISOLATION OF KERATINOPHILIC FUNGI FROM THE

FLOORS OF PRIVATE VETERINARY CLINICS IN ITALY

F. MANCIANTI’ AND R. PAPIN12

‘Dipartimento di Patologia Animale, Profilassi e Igiene degli Alimenti; ‘Istituto di
Patologia Speciale e Clinica Medica Veterinaria, Facolta di Medicina Veterinaria, Viale
delle Piagge 2, 56123 Pisa, Italy

ABSTRACT

Mancianti, F. and Papini, R., 1996. Isolation of keratinophilic fungi from the floors of private veterinary
clinics in Italy. Veterinary Research Communications, u)(2), 161-166

To evaluate the presence of keratinophilic fungi in the environment, 400 samples were collected from
the floors of 50 private veterinary clinics using 5%mm-diameter ‘contact plates’, containing mycobiotic
agar. After incubation for 15 days at 25”C, the following species were isolated: Microsporum canis,
Trichophyton terrestre, Chrysosporium keratinophilum, Chrysosporium sp., Microsporum gypseum,
Trichophyton ajelloi, Chrysosporium tropicum, Trichophyton mentagrophytes, Chrysosporium state of
Arthroderma tuberculatum and Chrysosporium pa-rum. It is concluded that the keratinic material
shed by infected pets may contribute to the development and propagation of dermatophytes and related
fungi in veterinary clinics. Therefore, such veterinary clinics may represent sites where pets and humans
are exposed to risk of infection with keratinophilic fungi from the environment.

Keywords: Chrysosporium, dermatophyte, epidemiology, floor, fungi, Microsporum, Trichophyton,

veterinary clinic

INTRODUCTION

The dermatophytes are the aetiological agents of human and animal mycoses,
belonging to the large group of keratinophilic fungi. These have the biological ability
to metabolize the keratinaceous substances that constitute the external surface of the
skin (hair, nails, epidermis, corneal scales and feathers). Some of them utilize keratin in
a saprobiotic manner, metabolizing only inert keratinic fragments. In contrast, the
dermatophytes have developed the biochemical activity to invade and utilize the
keratin on living hosts, so becoming parasites. These include the genera Epidermo-
phyton, Microsporum and Zlichophyton. This capacity is shared by some species of the
genus Chrysosporium, which are also similar to several true dermatophytes in sexual
reproduction through the teleomorphic stage termed Arthroderma. A pathogenic role
for the Chrysosporium species cannot be excluded (Krempl-Lamprecht, 1965; Stillwell
ef al., 1984).
In the field of medical mycology, the pathology connected with dermatophytes and
related fungal infections assumes a particular significance owing to the relationship
between humans, domestic animals and keratinophils: several reports have shown that
high levels of animal and human density favour the presence of keratinophilic fungi in
soil (Marsella et al., 1985; Mercantini et al., 1978, 1989). Hence, it seems probable that

161
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the isolation of Epidermophyton, Microsporum, Trichophyton and Chrysosporium

species from the environment is associated with the presence of keratinaceous material
shed by humans and animals frequenting particular places.
The present study aimed to verify this tendency of dermatophytes and related fungi
to be associated with the presence of animals by studying their presence on the floors of
private veterinary clinics in Italy.

MATERIAL AND METHODS

Samples were collected from floors of 50 private veterinary clinics for small animals in
some provinces of Tuscany. ‘Contact plates’ of 55 mm diameter (RODAC, PBI
International S.p.A., Milano, Italy) filled with 16.5 ml of mycobiotic agar (Difco
Laboratories, Chicago, IL, USA) were used. Two plates were applied, four times each,
directly onto randomly chosen sites on the floors of waiting rooms, examination
rooms, radiography rooms and wards. About 200 cm* of the surface of the floor was
tested in each room. Eight plates were collected on the same day from each veterinary
clinic, the total number obtained being 400 over a 15-day period. The samples were
always taken at the end of the working day, before the floors were cleaned, and were
cultured at 25°C for 15 days, being examined daily from the fourth day. Colonies of the
fungi so isolated were transferred onto Sabouraud dextrose agar (Difco Laboratories)
for identification based upon their macroscopical and microscopical features (Rippon,
1988).

RESULTS

The 10 species of keratinophilic fungi which were identified and the frequencies at
which they were isolated are shown in Table I.

TABLE I
Speciesof keratinophilic fungi isolated from the floors of veterinary clinics in Italy

Speciesisolated No. of positive No. of positive

veterinary clinics plates

Microsporum canis 15 46
Wchophyton terrestre 11 12
Chrysosporium keratinophilum 9 12
Chrysosporium sp. 5 5
Microsporum gypseum 5 6
Trichophyton ajelloi 4 5
Chrysosporium tropicum 4 7
Trichophyton mentagrophytes 2 2
Chrysosporium state of Arthroderma tuberculatum 2 3
Chrysosporium pannorum 1 2
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TABLE II
Associations of speciesof keratinophilic fungi isolated from the floors of veterinary clinics in
Italy

Speciesand mixed combinations of keratinophilic No. of positive Total no. of

fungi recovered (number of clinics) plates per clinic veterinary
clinics

Nil 0 12

M. canis (2); M. gypseum (2); I: menzugrophytes(1); 1 17

M. cunis + C. kerutinophilum (1); lY terrestre (3);
T. ujelloi (2); Chrysosporium sp. (2);
C. kerutitwphilum (3); C. kerutinophilum + C. tropicurn (1)

M. gypseum (1); Z ujelloi + Chrysosporium sp. (1);

T terrestre + C.A. tuberculutum (1);
C.A. tuberculutum (1); C. tropicum (2); C. punnorum (1)

M cunis (1); M. cunis + ‘I: terrestre (2);

M. cunis + lY mentugrophytes (1);
M. cunis, 27 terrestre + C. tropicum (1);
T. terrestre, Chrysosporium sp. + C. kerutinophdum (1)

M. cuds, M, gypseum, lY terrestre + C. kerutinophilum (1); 4 3

M. cunis + C. kerutinophilum (1); M. gypseum,
T. terrestre, C. kerutinophilum + Chrysosporium sp. (1)

M. eunis (1); M. cur&, I: terrestre + T ujefioi (1) 5 2

M. cunis (1) 6 1

M. cunis (1) 7 1

M. cunis (1) 8 1

6The occurrence of keratinophilic fungi varied considerably among the clinics: all the
plates gave negative results in 12 clinics. The frequencies of keratinophilic fungi and
mixed cultures are reported in Table II. Also, the occurrence of keratinophilic fungi
was not uniform among the tested rooms: 4 rooms were contaminated in 3 veterinary
clinics, 3 in 5, 2 in 9, and 1 in 21. The examination rooms were most frequently
contaminated (23/50), followed by the waiting rooms (18/50), the wards (14/50) and
the radiography rooms (11150). In most cases only one species was recovered from a
room but sometimes (15 / 50) two species were isolated.
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DISCUSSION

The results showed that veterinary clinics may be highly contaminated with dermato-
phytes and related fungi. Some of the species recorded in this study, including A4.
gypseum, 27 ajelloi, T. terrestre and the Chrysosporium species, are isolated from soils
where keratinaceous material accumulates (Otcenasek and Dvorak, 1964; Mercantini
et al, 1978, 1989) and may have been carried into the clinics on fomites (boots, shoes
etc.). These fungi show different pathogenic features; all possess the ability to
metabolize keratin but, while M. gypseum is a true geophilic dermatophyte, the role
of the other species is not completely clear. 57 ajelloi was isolated from tinea corporis in
a child (Presbury and Young, 1978) and from infections in cattle, dogs, horses and
squirrels (Rippon, 1988). i7 terrestre and some Chrysosporium species have been
obtained from the skin of small mammals (Marples and Smith, 1962; Chabasse et al.,
1987).
Some pathogenic zoophilic species were also isolated, including M canis and r
mentagrophytes. Their presence in the clinics is probably due to the introduction of
dogs and cats which were shedding infected keratinaceous material. It is known that
keratinophilic fungi in the environment can cause infections (Alexander et al., 1965;
Fujhiro, 1994; Thomas et al., 1994). Dermatophytes may remain viable in infected
material for many years, although the survival time depends on many factors,
including the nature of the infected material and the species of dermatophyte.
Arthrospores contained within hair, skin or crusts are protected from the lethal effects
of ultraviolet radiation and these materials are probably the major source of persistent
environmental contamination. Hence, keratinaceous debris shed by attending pets may
contribute to the propagation and development of dermatophytes in veterinary clinics
and represent a source of infection for other animals and humans. Their persistence in
the environment will be dependent on the quality and quantity of cleaning, and some of
the differences among the clinics may have been due to the efficacy of these procedures
rather than to the health status of the attending pets.
M. canis was the most interesting species isolated. This dermatophyte is the main
aetiological agent of ringworm in cats and dogs, and can affect man. Its prevalence in
animals and humans is high and it has also been isolated from environments where
humans are concentrated, such as schools, ferry-boats and trains (Mercantini et al.,
1986, 1989). In our investigations, M. canis was often isolated in pure culture.
Although records were not available of the number of M. can&infected cats and dogs
seen in the clinic just before the environmental examinations were carried out, we are
inclined to believe that its occurrence might be due to many cases of acute M. canis
infections occurring during the study, so that the arthrospores in the environment were
particularly abundant and viable. Contamination of the environment from clinical
cases may be extensive and there is evidence that this may be more severe in cases of IV.
canis than with other dermatophytes (Thomas et al., 1994).
In general, it is difficult to compare our results with those of other authors: most of
the latter’s studies have been carried out in sites with high human densities and this
may account for some differences encountered in the species isolated. We cannot
exclude the possibility that some microorganisms recovered from the floors were shed
 165

by people. However, strictly anthropophilic fungi, such as EpidermophytonJloccosum,

which was found in samples from schools, ferry-boats and trains (Mercantini et al.,
1986, 1989) were not recovered in the present investigation. Further differences may be
accounted for by the methods used. Previous workers employed a hair-bait method or
distribution of dust suspensions on selective cultural media, while in our study contact
plates were used. This method was chosen because it is effective, simple and stable.
Such plates can also be employed in quantitative analyses (Vanbreuseghem et al., 1979).
Mercantini and colleagues (1978) investigated the keratinophilic flora from the
floors of cages and animal enclosures of the zoo at Pescasseroli: keratinophilic species
were present in 100% of the samples, M. gypseum being the sole dermatophyte isolated
from 20% of the samples. In our investigation, M. canis was the most widespread
species (11.5%) and other keratinophilic fungi were present in 13% of samples taken.
These differences may be related to various factors such as the species of animals
examined, their health status, the type of flooring, the quality and frequency of
cleaning, and the quantity of keratinaceous material shed by the animals.
It is concluded that veterinary clinics should be considered to be sites where the
input of keratinophilic fungi is high, so that animals and humans may be exposed to
risk of infection with dermatophytes and related fungi from the environment. The
prolonged survival time of dermatophytes in clinical material is of considerable
epidemiological importance. Although direct contact may be a more efficient means
of transmission of dermatophytosis, indirect transmission through fomites and
environmental contamination can occur readily and may be more common. Since the
purpose of our study was to point out environmental contamination as a possible
source of keratinophilic fungi in veterinary clinics, the contact plates were not taken to
confirm the efficacy of the cleaning in removing infected hairs and scales. However, in
our opinion, it is advisable that removal of keratinic debris and disinfection (Rycroft
and McLay, 1991) specifically directed at limiting such a risk are regularly and
frequently undertaken.

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(Accepted: 22 September 1995)