AAC Accepted Manuscript Posted Online 20 April 2015 Antimicrob. Agents Chemother. doi:10.1128/AAC.00244-15 Copyright © 2015, American Society for Microbiology. All Rights Reserved.
1
Comparison of in vitro susceptibility of newer triazoles and established
2
antifungal agents against Trichophyton rubrum.
3 4
Shuwen Deng 1*, Chao Zhang 1*, Seyedmojtaba Seyedmousavi
5
Zhu 1,Xin Tan 5, Yiyang Wen 6, Xin Huang
6
Wenjie Fang 1, Shuaishuai Shen 6, Danqi Deng 5, Weihua Pan
7
Liao 1#
6
2, 3, 4,
Shuang
, Wenzhi Lei 6, Zhaojing Zhou 1, 1#,
Wanqing
8 9
1 Shanghai
Institute of Medical Mycology, Changzheng Hospital, Second
10
Military Medical University, Shanghai, China
11
2 Department
12
Netherlands
13
3 Department
14
The Netherlands
15
4
16
Sari, Iran
17
5 Department
18
Medical University, Kunming, China
19
6 Department
20
School of Medicine, Shanghai, China
of Medical Microbiology, Radboudumc, Nijmegen, The
of Medical Microbiology and Infectious Diseases, ErasmusMC,
Invasive Fungi Research Center, Mazandaran University Medical Center,
of Dermatology, The second affiliated hospital of Kunming
of Dermatology, Shanghai Tongji Hospital, Tongji University
21 22
Running title: Antifungal susceptibility of Trichophyton rubrum 1
23
Key words: Trichophyton rubrum, dermatophytosis, antifungal susceptibility
24
testing
25 26
*Correspondence: Prof. Wanqing Liao
27
Shanghai Institute of Medical Mycology, Changzheng Hospital, Second Military
28
Medical University, Shanghai, China
29
Address: Fengyang Rd No.415, Shanghai, China, 200003
30
Tel: + 86 (21) 81885439
31
E-mail: [email protected]
32
Co - Corresponding: Dr. Weihua Pan, E-mail: [email protected]
33
*Shuwen Deng and *Chao Zhang contributed equally to this study.
34 35
111 clinical Trichophyton rubrum isolates were tested against 7 antifungal
36
agents. Geometric mean MIC of all isolates were in increasing order:
37
terbinafine ( 0.03 mg/L ), voriconazole ( 0.05 mg/L ), posaconazole ( 0.11
38
mg/L ), isavuconazole ( 0.13 mg/L ), itraconazole ( 0.26 mg/L ), griseofulvin
39
( 1.65 mg/L ), fluconazole ( 2.12 mg/L ).
40 41
Wordcount abstract: 56
42
Wordcount maintext: 992
43 44
Dermatophytosis caused by Trichophyton rubrum are the most common 2
45
cutaneous fungal infections worldwide(1), which represents the cause of
46
between 80% and 90% of all chronic and recurrent infections(2). These
47
infections establish an important public health problem because of prolonged
48
treatment of the disease, usual recurrence infection are generally considered
49
difficult to manage(3). Reliable in vitro susceptibility testing would therefore be
50
useful for selecting the most suitable antifungal treatment. For many years,
51
griseofulvin was the only approved systemic anti-dermatophytic agent(4).
52
However, nowadays many potent antifungal agents are available for the
53
treatment of dermatophytosis, such as allylamines, and triazoles with the more
54
potent activity and less side effects(5-19). Expansion of information on in vitro
55
susceptibility testing of dermatophytes to new antifungal agents will help in
56
selecting or developing antifungal drug regimens.
57
The aim of the study was to compare in vitro the susceptibility of three newer
58
triazoles including voriconazole, posaconazole and isavuconazole and four
59
established antifungal agents against. T. rubrum. 111 clinical isolates of T.
60
rubrum were collected from seven dermatology clinics in Shanghai, China.
61
Morphological identifications were confirmed by sequence-based analysis of
62
the internal transcribed spacer of the rDNA region. In vitro activity of seven
63
antifungal agents was determined with the reference guideline M38-A2 (20)
64
with minor modification. Two reference strains, Trichophyton mentagrophytes
65
(ATCC-MYA-4439) and Candida parapsilosis ( ATCC 22019 ) were included
66
as quality controls. Student’s t test with the statistical SPSS package (version 3
67
9.0) were used and P values of 0.05 were considered statistically significant.
68
Table 1 listed the results of the MIC ranges, geometric mean MICs, MIC50 and
69
MIC90 values of seven antifungal agents against 111 T. rubrum strains.
70
Terbinafine,
71
griseofulvin, had low MICs, against all tested strains, whereas fluconazole did
72
not show inhibitory effects. Similarly, it has been shown in several studies
73
(table 2), however, limited data are available for newer triazoles isavuconazole,
74
posaconazole.
75
Terbinafine was one of the most effective antifungal agents against T.rubrum
76
among the 7 fungal agents tested and our findings confirmed those of previous
77
studies(5-19) (table 2).
78
We compared the in vitro activities of 3 newer triazoles isavuconazole,
79
posaconazole and voriconazole with itraconazole. Three newer triazoles
80
offered good in vitro activity against T. rubrum ( table 1 ). All isolates were far
81
more susceptible to the 3 newer triazoles than itraconazole (table 1), and
82
comparable to those reported by other studies(7, 9, 10, 14, 17, 18).
83
Isavuconazole is a novel broad-spectrum triazole agent and shares the same
84
mechanism of action with the other triazoles. Several studies supported its
85
efficacy in invasive Candida species, Cryptococcus neoformans, Aspergillus
86
species, Mucorales isolates(5-19, 21). However, the antifungal-susceptibility
87
profile of dermatophytes remains poorly examined. Ghannoum M et al.
88
reported that isavuconazole had shown potent in vitro activity against the
voriconazole,
posaconazole,
isavuconazole,
itraconazole,
4
89
dermatophytes(22) and was more active than other triazoles tested
90
(itroconazole, voriconazole ), but had higher MIC value than terbinafine,
91
however, against T. rubrum isolates with high MIC to terbinafine ,
92
isavuconazole remained low (0.06 mg/L for all tested isolates )(23). In our
93
study, MIC values of isavuconazole ( GM 0.13 mg/L, MIC90 0.125 mg/L ) were
94
similar to posaconazole (GM 0.11 mg/L, MIC90 0.5 mg/L ), and voriconazole
95
(GM 0.05 mg/L, MIC90 0.125 mg/L ), the difference was within 1 log2-dilution
96
step, much lower than itraconazole (GM 0.26 mg/L, MIC90 2 mg/L) for the
97
majority of T. rubrum tested.
98
Posaconazole showed similar activity as those published by A.K. Gupta who
99
reported posaconazole as the most active compound with MIC90 168
PDA
28
Adimi, P., et al. (2013)
M38-A2
0.007-0.5
0.04
0.031-1
0.1
-
-
-
-
M38-A
0.0156-16
0.172
0.0009-4
0.06
0.0078-8
0.19
0.0625-256
18.82
M38-A
0.0075-0.015
0.01
0.062-15
0.24
-
-
-
-
-
-
0.5-1
0.88
23
110
PDA
28
Ataides, F.S., et al.(2011)
M38-A
-
-
-
-
0.01-1
0.06
0.06-64
2.79
-
-
-
-
139
48,72,96,longer
PDA
28
Carrillo-Muñoz, A.J., et al.
M-38-A
0.001-0.03
0.006
0.25-2
0.059
-
-
0.5-64
1.92
0.06-1
0.21
-
-
16
110
PDA
30
Singh, J., M, et al.(2007)
M-38-A
8
0.07
-
-
0.125->64
5.36
0.007-0.5
-
-
73
168
OA
35
Gupta, A.K., et al.(2005)
M-27-A
0.003-1
0.003
0.06-32
0.14
-
-
-
-
-
-
-
68
168
OA
35
Gupta, A.K., et al.(2003)
M-38-P
16
0.01
0.018-1
0.09
0.01-1
0.06
0.06->64
2.8
-
-
-
-
144
96
PDA
28
M-27-A
0.01-0.25
0.01
0.03-1
0.08
1
Comparison of in vitro susceptibility of newer triazoles and established
2
antifungal agents against Trichophyton rubrum.
3 4
Shuwen Deng 1*, Chao Zhang 1*, Seyedmojtaba Seyedmousavi
5
Zhu 1,Xin Tan 5, Yiyang Wen 6, Xin Huang
6
Wenjie Fang 1, Shuaishuai Shen 6, Danqi Deng 5, Weihua Pan
7
Liao 1#
6
2, 3, 4,
Shuang
, Wenzhi Lei 6, Zhaojing Zhou 1, 1#,
Wanqing
8 9
1 Shanghai
Institute of Medical Mycology, Changzheng Hospital, Second
10
Military Medical University, Shanghai, China
11
2 Department
12
Netherlands
13
3 Department
14
The Netherlands
15
4
16
Sari, Iran
17
5 Department
18
Medical University, Kunming, China
19
6 Department
20
School of Medicine, Shanghai, China
of Medical Microbiology, Radboudumc, Nijmegen, The
of Medical Microbiology and Infectious Diseases, ErasmusMC,
Invasive Fungi Research Center, Mazandaran University Medical Center,
of Dermatology, The second affiliated hospital of Kunming
of Dermatology, Shanghai Tongji Hospital, Tongji University
21 22
Running title: Antifungal susceptibility of Trichophyton rubrum 1
23
Key words: Trichophyton rubrum, dermatophytosis, antifungal susceptibility
24
testing
25 26
*Correspondence: Prof. Wanqing Liao
27
Shanghai Institute of Medical Mycology, Changzheng Hospital, Second Military
28
Medical University, Shanghai, China
29
Address: Fengyang Rd No.415, Shanghai, China, 200003
30
Tel: + 86 (21) 81885439
31
E-mail: [email protected]
32
Co - Corresponding: Dr. Weihua Pan, E-mail: [email protected]
33
*Shuwen Deng and *Chao Zhang contributed equally to this study.
34 35
111 clinical Trichophyton rubrum isolates were tested against 7 antifungal
36
agents. Geometric mean MIC of all isolates were in increasing order:
37
terbinafine ( 0.03 mg/L ), voriconazole ( 0.05 mg/L ), posaconazole ( 0.11
38
mg/L ), isavuconazole ( 0.13 mg/L ), itraconazole ( 0.26 mg/L ), griseofulvin
39
( 1.65 mg/L ), fluconazole ( 2.12 mg/L ).
40 41
Wordcount abstract: 56
42
Wordcount maintext: 992
43 44
Dermatophytosis caused by Trichophyton rubrum are the most common 2
45
cutaneous fungal infections worldwide(1), which represents the cause of
46
between 80% and 90% of all chronic and recurrent infections(2). These
47
infections establish an important public health problem because of prolonged
48
treatment of the disease, usual recurrence infection are generally considered
49
difficult to manage(3). Reliable in vitro susceptibility testing would therefore be
50
useful for selecting the most suitable antifungal treatment. For many years,
51
griseofulvin was the only approved systemic anti-dermatophytic agent(4).
52
However, nowadays many potent antifungal agents are available for the
53
treatment of dermatophytosis, such as allylamines, and triazoles with the more
54
potent activity and less side effects(5-19). Expansion of information on in vitro
55
susceptibility testing of dermatophytes to new antifungal agents will help in
56
selecting or developing antifungal drug regimens.
57
The aim of the study was to compare in vitro the susceptibility of three newer
58
triazoles including voriconazole, posaconazole and isavuconazole and four
59
established antifungal agents against. T. rubrum. 111 clinical isolates of T.
60
rubrum were collected from seven dermatology clinics in Shanghai, China.
61
Morphological identifications were confirmed by sequence-based analysis of
62
the internal transcribed spacer of the rDNA region. In vitro activity of seven
63
antifungal agents was determined with the reference guideline M38-A2 (20)
64
with minor modification. Two reference strains, Trichophyton mentagrophytes
65
(ATCC-MYA-4439) and Candida parapsilosis ( ATCC 22019 ) were included
66
as quality controls. Student’s t test with the statistical SPSS package (version 3
67
9.0) were used and P values of 0.05 were considered statistically significant.
68
Table 1 listed the results of the MIC ranges, geometric mean MICs, MIC50 and
69
MIC90 values of seven antifungal agents against 111 T. rubrum strains.
70
Terbinafine,
71
griseofulvin, had low MICs, against all tested strains, whereas fluconazole did
72
not show inhibitory effects. Similarly, it has been shown in several studies
73
(table 2), however, limited data are available for newer triazoles isavuconazole,
74
posaconazole.
75
Terbinafine was one of the most effective antifungal agents against T.rubrum
76
among the 7 fungal agents tested and our findings confirmed those of previous
77
studies(5-19) (table 2).
78
We compared the in vitro activities of 3 newer triazoles isavuconazole,
79
posaconazole and voriconazole with itraconazole. Three newer triazoles
80
offered good in vitro activity against T. rubrum ( table 1 ). All isolates were far
81
more susceptible to the 3 newer triazoles than itraconazole (table 1), and
82
comparable to those reported by other studies(7, 9, 10, 14, 17, 18).
83
Isavuconazole is a novel broad-spectrum triazole agent and shares the same
84
mechanism of action with the other triazoles. Several studies supported its
85
efficacy in invasive Candida species, Cryptococcus neoformans, Aspergillus
86
species, Mucorales isolates(5-19, 21). However, the antifungal-susceptibility
87
profile of dermatophytes remains poorly examined. Ghannoum M et al.
88
reported that isavuconazole had shown potent in vitro activity against the
voriconazole,
posaconazole,
isavuconazole,
itraconazole,
4
89
dermatophytes(22) and was more active than other triazoles tested
90
(itroconazole, voriconazole ), but had higher MIC value than terbinafine,
91
however, against T. rubrum isolates with high MIC to terbinafine ,
92
isavuconazole remained low (0.06 mg/L for all tested isolates )(23). In our
93
study, MIC values of isavuconazole ( GM 0.13 mg/L, MIC90 0.125 mg/L ) were
94
similar to posaconazole (GM 0.11 mg/L, MIC90 0.5 mg/L ), and voriconazole
95
(GM 0.05 mg/L, MIC90 0.125 mg/L ), the difference was within 1 log2-dilution
96
step, much lower than itraconazole (GM 0.26 mg/L, MIC90 2 mg/L) for the
97
majority of T. rubrum tested.
98
Posaconazole showed similar activity as those published by A.K. Gupta who
99
reported posaconazole as the most active compound with MIC90 168
PDA
28
Adimi, P., et al. (2013)
M38-A2
0.007-0.5
0.04
0.031-1
0.1
-
-
-
-
M38-A
0.0156-16
0.172
0.0009-4
0.06
0.0078-8
0.19
0.0625-256
18.82
M38-A
0.0075-0.015
0.01
0.062-15
0.24
-
-
-
-
-
-
0.5-1
0.88
23
110
PDA
28
Ataides, F.S., et al.(2011)
M38-A
-
-
-
-
0.01-1
0.06
0.06-64
2.79
-
-
-
-
139
48,72,96,longer
PDA
28
Carrillo-Muñoz, A.J., et al.
M-38-A
0.001-0.03
0.006
0.25-2
0.059
-
-
0.5-64
1.92
0.06-1
0.21
-
-
16
110
PDA
30
Singh, J., M, et al.(2007)
M-38-A
8
0.07
-
-
0.125->64
5.36
0.007-0.5
-
-
73
168
OA
35
Gupta, A.K., et al.(2005)
M-27-A
0.003-1
0.003
0.06-32
0.14
-
-
-
-
-
-
-
68
168
OA
35
Gupta, A.K., et al.(2003)
M-38-P
16
0.01
0.018-1
0.09
0.01-1
0.06
0.06->64
2.8
-
-
-
-
144
96
PDA
28
M-27-A
0.01-0.25
0.01
0.03-1
0.08
