Medical Mycology 2002, 40, 201–208 Accepted 19 September 2001

Fungicidal activities of commonly used disinfectants and

antifungal pharmaceutical spray preparations against
clinical strains of Aspergillus and Candida species
A. K. GUPTA*, I. AHMADy & R. C. SUMMERBELLz
*
Division of Dermatology, Department of Medicine, Sunnybrook and Women©s College Health Sciences Center (Sunnybrook site),

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and the University of Toronto, Toronto, Ontario, Canada; yMedical Mycology, Laboratories Branch, Ontario Ministry of Health,
Toronto, Ontario, Canada and Mediprobe Laboratories, Toronto, Ontario, Canada; zDepartment of Laboratory Medicine and
Pathobiology, University of Toronto, Toronto, Ontario, Canada and Centraalbureau voor Schimmelcultures, Baarn, Netherlands

The antifungal efŽcacy of commercial chemical disinfectants and pharmaceutical

antifungal agents against medically important moulds and yeast species was
investigated. Chlorine, phenol, sodium dodecyl sulfate and quaternary ammonium
salts were the chemical disinfectants, and bifonazole and terbinaŽne were the
antifungal pharmaceutical products tested against clinical isolates of Aspergillus and
Candida species. Fungal inocula were obtained from conidial preparations of two
A. ochraceus strains and yeast cells of C. albicans, C. krusei and C. parapsilosis. The
antifungal activities were evaluated either by determining the kill rate in a cell
suspension media at different contact periods, or by examining the viability and
growth on plates sprayed with the active ingredient. Chlorine (1%) was the only
disinfectant with the ability to cause a rapid inactivation of all Žve strains. Phenol
(5%) was equally effective against Candida species; however, a number of
A. ochraceus conidia were able to survive this treatment for up to 1 h.
Benzalkonium chloride (0 5%) and cetrimide (0 5%) were also able to disinfect
the three Candida species rapidly; however, these two quaternary ammonium
compounds were relatively ineffective against A. ochraceus. In spray experiments,
quaternary ammonium compounds had a fungicidal activity against Candida species
and were fungistatic against A. ochraceus conidia. All Žve fungal strains were able to
resist 0 5% sodium dodecyl sulfate, present either in the suspension solution or on
the sprayed plate. Of the two pharmaceutical antifungal products tested, bifonazole
(1%) were essentially ineffective against all Žve strains. TerbinaŽne (1%) had a
fungicidal activity against A. ochraceus and C. parapsilosis. In suspension experi-
ments, an exposure to 0 01% terbinaŽne required a contact period of 1 h for a
complete inactivation of A. ochraceus conidia and an onset of fungicidal effect on
C. parapsilosis yeast cells. TerbinaŽne was only moderately effective against
C. albicans and was completely ineffective against C. krusei.
Keywords antifungal drugs, Aspergillus ochraceus, Candida species, chemical
disinfectants, fungicidal, fungistatic, nondermatophytes

Correspondence: Aditya K. Gupta M.D., Ph.D. F.R.C.P.(C), 490

Wonderland Road South, Suite 6, London, Ontario, Canada N6K
1L6. Tel.: ‡1 519 657 4222; Fax: ‡1 519 657 4233; E-mail: agupta@
execulink.com.
ÓÓ 2002
2002 ISHAM
ISHAM, Medical Mycology, 40, 201–208
 202 Gupta et al.
Introduction
Pathogenic strains of Candida and Aspergillus species
are the primary cause of hospital-acquired fungal
infections [1], and in recent years these yeast and mould
infections have been recognized as serious threats to
immunocompromised patients [2–4]. The distribution of
these fungal pathogens appears to be widespread in our
environment. In dermatological laboratory examina-
tions, Candida and Aspergillus species are frequently
isolated as superŽcial infections of skin and nails [5,6].
The shedding of dermatophytes and other fungal
pathogens from infected individuals often leads to the
contamination of recreation facilities in public arenas [7],
and the eradication of this contamination from public
facilities such as swimming pools has been found to be an
arduous task [8]. These facts underscore the importance
of Žnding adequate measures for eradicating fungal
contaminants in our immediate environment. Several
new disinfectants containing quaternary ammonium
compounds as active agents have been introduced in
the market in recent years. Many of these formulations
are stated to have a broadly based antifungal activity;
however, the evidence for their efŽcacy against medi-
cally important fungi is not clear in the literature. The
work of Ohta et al. [9] indicates a considerable variation
in the ability of commercially available quaternary
ammonium compounds to disinfect clinical strains
belonging to Aspergillus and Candida species. In the
US, all the Environmental Protection Agency (EPA)-
registered quaternary-ammonium-compound-based anti-
fungal formulations failed the disinfectant test against
clinical strains of Aspergillus species [10]. These studies
indicate some serious shortcomings in the efŽcacy of
prevalent antifungal commercial disinfectants, and there
appears to be a need for new approaches to disinfecting
pathogenic fungi in our environment. The work of
Piérard et al. [11] with miconazole powder suggest that
pharmaceutical products can be employed superŽcially
as protective agents against infectious fungi present on
external surfaces. Fortunately, a number of antifungal
Table 1 Clinical isolates
Vegetative
Species morphology
A. ochraceus Filamentous
A. ochraceus Filamentous
C. albicans Yeast
C. krusei Yeast
C. parapsilosis Yeast
*Ontario Ministry of Health laboratory specimen accession number.
pharmaceutical products have become available in spray
formulations in recent years. It is pertinent, therefore, to
evaluate the efŽcacy of antifungal drugs to disinfect
fungal pathogens prevalent in our environment. Cur-
rently, the disinfection of various groups of medically
important fungi is being investigated in our laboratory.
Our work on the disinfection of dermatophytic species of
Trichophyton has been presented previously [12]. The
present paper deals with the disinfection of Aspergillus
and Candida species. The antifungal activities of several
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commonly used chemical disinfectants and two recently
introduced pharmaceutical antifungal sprays has been
evaluated against standardized inocula of A. ochraceus,
C. albicans, C. krusei and C. parapsilosis.
Materials and methods
Clinical isolates
Freshly isolated strains of Aspergillus and Candida
species were obtained from clinical material at Labora-
tories Branch of Ontario Ministry of Health. Table 1 lists
two isolates of A. ochraceus and single isolates of
C. albicans, C. krusei and C. parapsilosis employed in
the present study. Cultures were maintained at 28 oC by
monthly transfer on Sabouraud’s agar medium (10 g l¡1
of Bacto peptone ‡ 40 g l¡1 of glucose ‡ 15 g l¡1 of
Difco agar). For experimental work, Candida yeast cells
were obtained from 1-week-old fungal lawns grown on
15 ml of Sabouraud’s agar in a 8 5-cm diameter petri
dish. Conidia of A. ochraceus were obtained from 2–3-
week-old fungal lawns grown on 80 ml of Sabouraud’s
agar in a 15-cm diameter petri dish.
Preparation of fungal inocula
The preparation of fungal inocula was carried out under
axenic conditions. Conidial cultures of A. ochraceus
were ooded with 20 ml of phosphate-buffered saline
(PBS), pH 7 2, gently scraped with a glass rod and the
resulting conidial suspension was gradually loaded with a
Pasteur pipette on a 5-ml plastic column padded with a
Designated OMH* Inoculum cell density
number (cfu ml¡1 )
FR-01382 47 106
SF-06430 36 106
FR-00806 37 108
FR-01190 63 108
FR-01760 55 108
Ó 2002 ISHAM, Medical Mycology, 40, 201–208
 Disinfectants and antifungals against Aspergillus and Candida 203

porous disk. Any hyphal debris on the top of the disk was Measurement of kill rates
cleared with a metal loop. The Žltered conidial suspen-
The kill rate of each disinfectant solution was deter-
sion was collected in a centrifuge tube and made up to
mined by examining the viability of fungal cells after 0-,
20 ml with PBS, pH 7 2. Candida cultures were scraped
0 25-, 0 5-, 1 0- and 24-h periods of contact with the
off gently with a heat-sterile scalpel and suspended in
disinfectant solution. The treatment was initiated by
20 ml of PBS, pH 7 2. These Candida preparations
adding 0 5 ml of quantiŽed fungal inoculum to 4 5 ml of
yielded homogenous turbid suspensions of yeast cells
disinfectant solution and was terminated by centrifuging
without any evidence of clumps or debris under
the suspension at 2000g for 5 min at room temperature
microscopic examinations. Therefore, Žltration was not
and suspending the fungal pellet in 10 ml of fresh PBS,
considered necessary for these preparations. For cell

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pH 7 2. To examine the presence of living cells in these
density measurements, each preparation was serially
suspensions, a tenfold dilution of A. ochraceus and a
diluted and at each dilution triplicate 100-m l fractions
1000-fold dilution of Candida suspensions were pre-
were inoculated on 8 5-cm Sabouraud’s agar plates. The
pared, and triplicate 100-m l fractions were inoculated on
inoculum was spread evenly on the plate surface with a
10-ml Sabouraud’s slants in 2 5 5cm screw-capped
metal loop. The cell density of the inoculum was
bottles. The inocula were spread evenly on agar slants
estimated at the dilution yielding 100–300 individual
with a metal loop and the slants were incubated
colonies per culture plate. The cell density values
horizontally at room temperature. The slants were
presented in Table 1 correspond to the 20-ml inoculum
examined regularly for the emergence of fungal colonies.
prepared for each strain. The disinfection studies were
For each contact time, the highest colony count obtained
initiated within 2 h of inocula preparation.
over a 7-day growth period was recorded as cfus.

Preparation of disinfectant suspension solutions

Stock solutions of 10% chlorine (sodium hypochlorite),
50% phenol, 5% benzalkonium chloride (BAC), 5%
hexadecyltrimethyl ammonium bromide (cetrimide), 5%
sodium dodecyl sulfate (SDS) were prepared in auto-
claved distilled water. All reagents used were of
analytical grade. TerbinaŽ ne stock was prepared by
1 1
extracting a powdered 250-mg Sandoz Lamisil tablet
(Sandoz Pharmaceutical Corporation, East Hanover, NJ,
USA) for 30 min in 2 5 ml of dimethyl sulfoxide,
clarifying the extract by centrifugation at 1000g for
5 min and serially diluting the clariŽed extract in 80%
ethanol to obtain a 0 1% stock solution. All stock
solutions were stored at room temperature. Disinfectant
suspension solutions were prepared by adding 0 5 ml of
stock solution to 4 ml of autoclaved PBS, pH 7 2.
Negative controls were prepared by adding 0 5 ml of
autoclaved water to 4 ml of PBS, pH 7 2.
Disinfectant spray formulations
Disinfectant solutions of 5% phenol, 0 5% BAC, 0 5%
cetrimide were prepared in autoclaved water from stock
solutions and transferred to sterile 500-ml amber bottles
Žtted with plastic spray assemblies. The spray bottle
produced 0 6 ml of Žne mist in each delivery. Commer-
cially available brands of Lysol domestic spray (Reckitt
& Coleman Inc., Toronto, Ontario, Canada), Spray
1
Nine (Korkay System Canada Ltd., Gananoque, On-
tario, Canada) and Brentdale germicidal spray (Brent-
dale Chemicals, Rexdale, Ontario, Canada), each
containing a speciŽc formulation of quaternary ammo-
nium compounds (Table 2), were obtained locally.
1
Mycospor spray containing 1% bifonazole (Bayer
1
AG, Germany) and Lamisil spray containing 1%
terbinaŽne–HCl (Novartis, Spartan, Kempton Park,
South Africa) were obtained as physician samples.

Table 2 Antifungal spray formulations

Trade name Active ingredients

1
Lysol 0 1% n-Alkyl (40% C1 2 , 50% C14 , 30% C1 6 ) dimethylbenzylammonium chloride
1
Spray Nine 0 15% n-Alkyl (5% C1 2 , 60% C14 , 30% C1 6, 5% C18 ) dimethylbenzylammonium chloride
0 15% n-alkyl (68% C12 , 32% C1 4 ) dimethylethylbenzylammoniumchloride
Brentdale 0 1% n-Alkyl (5% C12 , 60% C1 4 , 30% C1 6, 5% C1 8 ) dimethylbenzylammonium chloride
0 1% n-Alkyl (68% C1 2 , 32% C14 ) dimethylethylbenzylammonium chloride
1 6% Tetrasodium ethylenediamine tetraacetate
1
Mycospor 1% Bifonazole
1
Lamisil 1% TerbinaŽne hydrochloride

Ó 2002 ISHAM, Medical Mycology, 40, 201–208

 204 Gupta et al.

Spray experiments for disinfectant tests within 3 months of its isolation.

Over this period, the subcultures of both Aspergillus
For each spray treatment, triplicate 100-m l aliquots of
strains and three Candida species maintained their
quantiŽ ed inoculum were evenly plated on 15 ml of
normal colony appearance, thereby ruling out any
Sabouraud’s agar medium in 8 5-cm diameter petri
apparent deterioration of these isolates on the growth
dishes and left for 2 h at room temperature. Plates were
media. In preliminary studies, a 24-h suspension of the
treated with the given disinfectant by spraying a thin Žlm
fungal inoculum in PBS, pH 7 2, followed by a 5-min
of about 1 8 0 2 ml on the agar surface. This layer of
centrifugation at 2000g had no detectable effect on the
spray on the agar plate covered with an unsealed lid
viability of A. ochraceus conidia and Candida yeast cells
evaporated completely within 3 days of incubation at
(results not shown).

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room temperature. On day 3, a surface swab from of
The effect of each disinfectant on the Žve fungal
each plate was inoculated onto a 10-ml Sabouraud’s slant
strains was evaluated either by determining the number
in a 2 5 5-cm bottle. Both sprayed plates and swab test
of cells surviving after a given period of cell contact with
slants were examined regularly, and the numbers of
the disinfectant in a suspension solution (Tables 3 and 4),
newly emerging colonies were recorded for a period of
or by counting the number of colonies able to grow on
14 days. Any spray formulation allowing more than
agar plates sprayed with the disinfectant (Tables 5 and
300 fungal colonies on the sprayed agar plate was
6). The cfus presented in Tables 3 and 4 correspond to
categorized as noninhibitory. The spray was categorized
100 m l of an appropriately diluted cell suspension that
as moderately inhibitory when the number of colonies on
was inoculated on a 10-ml agar slant in a 16-ml tube.
the plate ranged between 100 and 300. A fungistatic
Under these conditions, the water controls for each of
action was recorded when a large area of the sprayed
the Žve fungal strains produced a dense growth with
plate was essentially free of fungal growth, whereas the
more than 100 overlapping colonies covering the entire
swab test from the colony-free plate surface was positive
agar slant. Any other treatment allowing normal growth
for fungal growth. The spray was categorized as
with more than 100 colonies on the agar slant was,
fungicidal when the sprayed plate was essentially free
therefore, evaluated as noninhibitory. Similarly, in spray
of fungal growth and the swab test was negative for
experiments (Tables 5 and 6), water controls produced a
growth.
dense growth of more than 300 individual colonies on
the 8 5-cm diameter agar plate, and therefore, any other
Results
spray treatment allowing the growth of more than
Table 1 lists the two Žlamentous strains of A. ochraceus 300 colonies on the plate was considered noninhibitory.
and the three yeast species of Candida grown from In A. ochraceus, the inactivation of conidia occurred
primary cultures of clinical isolates. Each isolate was most rapidly in the presence of chlorine (Table 3). Both
transferred to fresh agar media every 4 weeks and used FR-01382 and SF-0643 were killed within 15 min of

Table 3 Inactivation of A. ochraceus conidia in disinfectant suspension solutions

Contact time

Treatment Isolate no. 15 min 30 min 60 min 24 h

Water FR-01382 > 100 > 100 > 100 > 100
SF-06430 > 100 > 100 > 100 > 100
1% Chlorine FR-01382 0 0 0 0
SF-06430 0 0 0 0
5% Phenol FR-01382 18 7 9 3 6 3 0
SF-06430 10 3 6 2 4 2 0
0 5 % BAC FR-01382 > 100 > 100 64 17 51 24
SF-06430 > 100 > 100 95 22 73 18
0 5% Cetrimide FR-01382 > 100 > 100 85 28 80 19
SF-06430 > 100 > 100 > 100 75 15
0 5% SDS FR-01382 > 100 > 100 > 100 > 100
SF-06430 > 100 > 100 > 100 > 100
0 01% TerbinaŽne FR-01382 6 2 7 3 2 1 0
SF-06430 19 4 6 3 8 3 0

Values are averages of three agar slants (cfus) standard deviations.

Ó 2002 ISHAM, Medical Mycology, 40, 201–208

 Disinfectants and antifungals against Aspergillus and Candida 205

Table 4 Inactivation of C. albicans FR-00806, C. krusei FR-01190 and C. parapsilosis FR-01760 in

disinfectant suspension solutions

Contact time

Treatment Species 15 min 30 min 60 min 24 h

C. albicans > 100 > 100 > 100 > 100

Water C. krusei > 100 > 100 > 100 > 100
C. parapsilosis > 100 > 100 > 100 > 100
C. albicans 0 0 0 0

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1% Chlorine C. krusei 0 0 0 0
C. parapsilosis 0 0 0 0
C. albicans 0 0 0 0
5% Phenol C. krusei 0 0 0 0
C. parapsilosis 0 0 0 0
C. albicans 8 3 5 2 0 0
0 5 % BAC C. krusei 0 0 0 0
C. parapsilosis 0 0 0 0
C. albicans 4 2 3 1 0 0
0 5% Cetrimide C. krusei 0 0 0 0
C. parapsilosis 14 5 10 4 0 0
C. albicans > 100 > 100 > 100 > 100
0 5% SDS C. krusei > 100 > 100 > 100 87 22
C. parapsilosis > 100 > 100 > 100 > 100
C. albicans > 100 > 100 > 100 85 22
0 01% TerbinaŽne C. krusei > 100 > 100 > 100 > 100
C. parapsilosis > 100 > 100 > 100 56 14

Values are averages of three agar slants (cfus) standard deviations.

contact with 1% chlorine in the suspension solution.
With 5% phenol, a number of conidia were able to
survive a contact of up to 60 min in the suspension
media. However, a 24-h contact with 5% phenol in the
suspension media and a 3-day incubation on phenol
sprayed agar plates was found to be fungicidal against
both strains of A. ochraceus (Tables 3 and 5). With 0 5%
BAC or cetrimide in the suspension media, no inactiva-
tion of A. ochraceus was detectable for a period of
30 min, and even after 24 h of contact a large number of
conidia were able to remain viable (Table 3). A 3-day
incubation with BAC and cetrimide on sprayed plates
caused a complete suppression of A. ochraceus conidia;
however, the swab tests from these plates revealed that
the effects of these two quaternary ammonium com-
pounds were largely fungistatic in nature. The three

Table 5 Inactivation of A. ochraceus by antifungal sprays

Isolate no.

FR-01382 SF-06430

ClassiŽcation of ClassiŽcation of
Spray cfus action cfus action

Water > 300 Noninhibitory > 300 Noninhibitory

5% Phenol 0 Fungicidal 0 Fungicidal
0 5 % BAC 0 Fungistatic 0 Fungistatic
0 5% Cetrimide 0 Fungistatic 3 2 Fungistatic
0 5% SDS > 300 Noninhibitory > 300 Noninhibitory
1
Lysol 0 Fungistatic 4 1 Fungistatic
1
Spray Nine 0 Fungistatic 0 Fungistatic
Brentdale 4 2 Fungistatic 0 Fungistatic
1
Mycospor > 300 Noninhibitory > 300 Noninhibitory
1
Lamisil 0 Fungicidal 0 Fungicidal

Values are averages of three agar plates standard deviations.

Ó 2002 ISHAM, Medical Mycology, 40, 201–208

 206 Gupta et al.

Table 6 Inactivation of C. albicans FR-00806, C. krusei FR-01190 and C. parapsilosis FR-01760 by antifungal sprays

C. albicans C. Krusei C. parapsilosis

ClassiŽcation of ClassiŽcation of ClassiŽcation of

cfus action cfus action cfus action

Water > 300 Noninhibitory > 300 Noninhibitory > 300 Noninhibitory
5% Phenol 0 Fungicidal 0 Fungicidal 0 Fungicidal
0 5 % BAC 0 Fungicidal 0 Fungicidal 0 Fungicidal
0 5% Cetrimide 0 Fungicidal 0 Fungicidal 0 Fungicidal

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0 5% SDS > 300 Noninhibitory > 300 Noninhibitory > 300 Noninhibitory
1
Lysol 0 Fungicidal 0 Fungicidal 0 Fungicidal
1
Spray Nine 0 Fungicidal 0 Fungicidal 0 Fungicidal
Brentdale 0 Fungicidal 0 Fungicidal 0 Fungicidal
1
Mycospor > 300 Noninhibitory > 300 Noninhibitory > 300 Noninhibitory
1
Lamisil 184 30 Inhibitory > 300 Noninhibitory 0 Fungicidal

Values are averages of three agar plates standard deviations.

commercial spray formulations of quaternary ammo-
1 1
nium compounds, Lysol , Spray Nine and Brentdale,
also produced a fungistatic effect against A. ochraceus
conidia on agar plates. SDS was completely ineffective
against A. ochraceus both in suspension media and on
sprayed plates (Tables 3 and 5). Of the two pharmaceu-
tical spray formulations used here, 1% bifonazole in
1
Mycospor was completely noninhibitory, whereas 1%
1
terbinaŽne in Lamisil spray was highly fungicidal
against this mould (Table 5). Even with a lower
concentration of 0 01% terbinaŽ ne in the suspension
media, only a few conidia of A. ochraceus were able to
survive the Žrst hour of contact with this drug (Table 3).
Both 1% chlorine and 5% phenol were fungicidal
within 15 min of contact with all three species of
Candida (Table 4). A 15-min contact with 0 5% BAC
was similarly lethal for C. krusei and C. parapsilosis
cells, whereas a contact period of 60 min was required
for a complete inactivation of C. albicans by this
compound (Table 4). Similarly, with 0 5% cetrimide in
the suspension media, C. krusei was killed within 15 min
of contact whereas a few cells of C. albicans and
C. parapsilosis survived for up to 60 min under these
conditions (Table 4). The three Candida species were
completely suppressed by BAC and cetrimide spray on
the agar plate, and the swab tests showed the inhibition
of Candida cells by the two quaternary ammonium
compounds to be fungicidal (Table 6). Quaternary
1 1
ammonium compounds in Lysol , Spray Nine and
Brentdale spray formulations were also fungicidal
against the three Candida species. The presence of
0 5% SDS, either in the suspension media (Table 4) or
on sprayed plates (Table 6), did not cause any inhibition
in the three Candida species. Spraying agar plates with
1
1% bifonazole (Mycospor ) was also noninhibitory to
the Candida species (Table 6). The response of the three
yeast species to the presence of terbinaŽ ne was, on the
other hand, quite variable. A spray with 1% terbinaŽ ne
had no effect on C. krusei, caused a moderate inhibition
in C. albicans, and brought about a fungicidal inactiva-
tion in C. parapsilosis. With 0 01% terbinaŽ ne in the
suspension media, a contact period of 24 h was required
to cause a detectable inactivation in C. albicans and
C. parapsilosis (Table 4).
Discussion
Disinfectants are deŽned as chemical agents capable of
removing infectious microbes other than bacterial spores
[13]. A number of commonly used disinfectants are
known to be relatively ineffective against fungi [14].
Moreover, not all fungal species are equally sensitive to a
given product, and even different strains of the same
fungal species may vary in resistance [14]. We have
monitored the ability of antimicrobial agents both in
suspension solutions and on sprayed culture plates. The
relevance of these two measures of antifungal activities
to the disinfection of various surfaces in our immediate
environment and the adequacy of the methodology used
is discussed elsewhere [12].
Chlorine was the only agent capable of killing within
15 min all Žve fungal strains tested in the present study.
Phenol was equally rapid in its fungicidal action against
the three Candida species; however, its action against the
two strains of A. ochraceus was considerably slower.
Both chlorine and phenol are highly toxic and corrosive
and their use has declined considerably in recent years
[14]. In today’s market, quaternary ammonium com-
pounds have become the most common ingredients of
disinfectant formulations. In our suspension experi-
ments, a contact of up to 24 h with BAC and cetrimide
caused only a partial inactivation of A. ochraceus
Ó 2002 ISHAM, Medical Mycology, 40, 201–208

conidia. A complete inactivation of A. ochraceus conidia
by BAC, cetrimide and the three commercial formula-
1
tions of quaternary ammonium compounds, Lysol ,
1
Spray Nine and Brentdale was observed after a 3-day
long exposure of these cells on sprayed plates. A high
level of resistance in A. ochraceus strains to chemical
disinfectants appears to be consistent with the data
published on clinical isolates of A. fumigatus, A. versi-
color, A. niger and A. terreus [9,10,16]. Quaternary
ammonium compounds do not appear to be sufŽciently
potent against clinical strains of Aspergillus species. SDS,
recently described [15] as an antimicrobial surfactant,
was ineffective against the Žve fungal strains tested in the
present study.
TerbinaŽ ne was notably fungicidal against both strains
of A. ochraceus. No viable conidia of the two strains
were found from agar plates sprayed with 1% terbina-
Žne. Even with a much lower concentration of 0 01%
terbinaŽne used in the suspension media, only a few
individual conidia remained viable after a 60-min
contact, and were completely inactivated after a 24-h
exposure to the drug. A concentration of 0 01%
terbinaŽne was formulated for our contact experiments,
because a similar level has been shown to separate
terbinaŽne sensitive and resistant cells in Candida
species [17].
The action of terbinaŽne against Candida species was
highly variable, with only C. parapsilosis showing a
marked sensitivity to the presence of this drug. Spraying
with 1% terbinaŽne resulted in a complete loss of
viability in C. parapsilosis. However, the action of
terbinaŽne on C. parapsilosis appears to be slow,
requiring a contact period of about 24 h with 0 01%
terbinaŽne for the onset of fungicidal activity. It may be
added here that for C. parapsilosis minimum inhibitory
concentrations (MICs) values of terbinaŽne are reported
to be in the range of 0 03-0 25 m g ml¡1 [17]. The
presence of 0 01% (100 m g ml1) terbinaŽ ne was, there-
fore, greatly in excess of that required to inhibit
C. parapsilosis cells, so the observed delay in terbinaŽ ne
action against this yeast species cannot be attributed to
the use of a suboptimal concentration of terbinaŽ ne in
our suspension media. The apparent lag in terbinaŽ ne
activity, therefore, appears to be an inherent phenom-
enon probably associated with the uptake rate and
accumulation of the drug by C. parapsilosis. The pattern
of resistance to terbinaŽ ne in Candida species –
C. krusei > C. albicans > C. parapsilosis – observed
in the present study agrees well with the data published
by Ryder et al. [17]. Bifonazole was relatively ineffective
in disinfecting the Žve fungal strains tested in the present
study.
The observed tolerance of Candida to terbinaŽne and
Ó 2002 ISHAM, Medical Mycology, 40, 201–208
Disinfectants and antifungals against Aspergillus and Candida 207
A. ochraceus to quaternary ammonium compounds
indicate the limitations in applying any given class of
antifungal agents as a common disinfectant against
medically important fungi. The feasibility of formulating
a combination of antimicrobial agents to target a broad
group of fungal pathogens needs to be tested. These
cocktails of antifungal agents could be a combination of
both chemical disinfectants and antifungal drugs.
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Acknowledgements
We thank Sal Albreish and his staff for a gift of
Aspergillus and Candida cultures.
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