JOURNAL OF CLINICAL MICROBIOLOGY, Nov. 1982, p. 976-978 Vol. 16, No.

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0095-1137/82/110976-03$02.00/0
Copyright © 1982, American Society for Microbiology

Nature and Identification of Exophiala werneckii

W. Y. MOK
Departamento de Patologia Tropical, Instituto Nacional de Pesquisas da AmazBnia, 69000 Manaus, Brazil
Received 22 April 1982/Accepted 26 July 1982

The morphological and physiological characteristics of 44 isolates of Exophiala

werneckii recovered from human and environmental sources were indistinguish-
able from 2 isolates that caused tinea nigra. Casein hydrolysis and inability to
decompose tyrosine differentiate E. werneckii from Exophiala jeanselmei, Exo-
phiala spinifera, and Wangiella dermatitidis.

Exophiala werneckii (Horta) Arx 1970 is the thermotolerance were studied by inoculating
etiological agent of the superficial mycosis tinea each isolate onto duplicate tubes of SAB agar
nigra (1, 4, 8, 10). Its distinguishing morphologi- and Czapek-Dox agar and incubating the cul-
cal feature is a yeast form which is typically two tures at 25, 28, 38, 42, and 45°C for 1 month.
celled. These yeast cells are annellides that form Weekly observations on colonial morphology
one- and two-celled conidia. The conidiogenesis were made.
and dimorphism of this fungus have been exam- Each isolate was inoculated onto duplicate
ined extensively (6, 7, 9, 13, 14). E. werneckii is tubes of SAB agar containing NaCl at 10, 20, and
differentiated from similar fungi solely by mor- 30% (wt/vol) at 25°C for 1 month. Weekly obser-
phology. The physiological properties of this vations were made. The biochemical properties
fungus have not yet been adequately studied. examined were hydrolysis of casein, tyrosine,
Recently, we isolated E. werneckii from sev- xanthine, hypoxanthine, and starch; gelatin liq-
eral unusual human and environmental sources. uefaction; paraffin utilization; sodium nitrate
One isolate was recovered from an inflammatory utilization in Czapek-Dox agar; proteolytic ac-
scalp lesion in a girl, and another came from a tivity in Loeffler coagulated serum; and fermen-
macerated interdigital lesion in a man's foot. tation of glucose, maltose, sucrose, galactose,
Two isolates were associated with plaquelike and lactose. The tests were performed by the
Candida infections in the crural areas of pa- methods described by Beneke and Rogers (2),
tients' bodies, and another was associated with a Berd (3), Conti-Diaz et al. (5), McClung (12),
case of tinea capitis already under treatment. In Silva (21), and Silva-Hutner and Cooper (22).
none of these five cases did we notice pigmented For comparison purposes, temperature and
macules at the infection sites or fungal elements biochemical studies were also carried out with
in epidermal scrapings. Additional isolates were the following fungi: Exophiala jeanselmei (Insti-
obtained from asymptomatic superficial human tuto de Higiene de Montevideo [IHM] isolates
dermal areas and from superficial moldy plaques 1860, 1783), Exophiala spinifera (IHM isolates
in spoiled salted freshwater fish (16). The objec- 1610, 1611, 1740, 1781), and Wangiella dermati-
tives of our study were to confirm the identity of tidis (IHM isolates 1763, 1765, 1766 and INPA
the E. werneckii isolates and to evaluate com- isolates 105, 109, 119, 122, 133, S11, S76, S93).
monly employed physiological tests as possible The IHM cultures had been examined and were
species-specific diagnostic criteria to supple- furnished by I. A. Conti-Diaz of the IHM, Mon-
ment the morphological identification of E. wer- tevideo, Uruguay (5). The INPA isolates were
neckii. recovered from bats (20) and soil (15) in Manaus,
We studied 46 isolates of E. werneckii: 5 were Brazil.
recovered from superficial dermal lesions in Colonies of the E. werneckii isolates on SAB
humans, 34 from asymptomatic human scalp as agar and Czapek-Dox agar were initially yeast-
well as the interdigital areas of hands and feet, 5 like and shiny black. With age, they produced
from moldy plaques on air-dried salted Amazo- submerged peripheral hyphae and abundant ae-
nian fish (Osteoglossum bicirrhosum), and 2 rial mycelia and turned olive in color. Colonial
controls- (B2803a, B2804a) from human tinea development was similar at 25, 28, and 38°C.
nigra. The 44 test isolates were maintained in the There was diminished metallic tonality in the
fungal culture collection at the Instituto Na- colonies of cultures kept at the higher tempera-
cional de Pesquisas da Amaz6nia (INPA). tures. At 42°C, all isolates showed slow but
All isolates were maintained on Sabouraud steady growth. At 45°C, there was no growth.
dextrose (SAB) agar. Colonial morphology and Microscopically, all isolates showed early hy-
976
VOL. 16, 1982
TABLE 1. Physiological characteristics of some Exophiala and Wangiella species
Fungus
No.
isolatesofDeopstoof
Decomposition of:
tested
Casein Tyro- Xan- Hypoxan-
sine thine thine Starch Protein faction tion
E. werneckii
Non-tineanigra 44 + - - -
isolates
Tinea nigra 2 + - -
isolates
E. jeanselmeia 2 - + -
E. spinifera 4 - + -
W. dermatitidis
IHM isolates 3 - + -
INPA isolates 8 - + -
a Classified as Phialophora gougerotii by the original authors (5).
b
Reported as 41°C by the original authors (5).
phal development and characteristic annellides
and annelloconidia, some being two celled with
prominent darkly pigmented septa. Chlamydo-
conidia and spiral hyphae were also present. All
structures were discernible as early as 10 days
after inoculation. There was no difference in
morphological development among the cultures
of the 44 test isolates and those of the 2 control
isolates on SAB agar, potato dextrose agar, and
cornmeal agar. However, growth was more rap-
id and abundant on cornmeal agar.
All E. werneckii isolates were resistant to high
salinity (30%) in the culture medium. The colo-
nial development of isolates on SAB agar satu-
rated with NaCl was indistinguishable from
those grown on SAB agar without NaCl. The
test and control isolates consistently decom-
posed casein but did not hydrolyze tyrosine,
xanthine, hypoxanthine, or starch. All isolates
utilized sodium nitrate in Czapek-Dox agar.
However, none liquefied gelatin or utilized par-
affin, except for two INPA isolates which
showed a very slight degree of paraffin utiliza-
tion. The isolates also did not hydrolyze protein
in Loeffler coagulated serum. None of the iso-
lates fermented glucose, maltose, sucrose, ga-
lactose, or lactose.
Table 1 presents data on the physiological
characteristics and maximum growth tempera-
tures of the E. werneckii, E. jeanselmei, E.
spinifera, and W. dermatitidis isolates. The fol-
lowing characteristics are being reported for the
first time: inability of E. spinifera to hydrolyze
xanthine, lack of paraffin utilization by E. spini-
fera and W. dermatitidis, and utilization of sodi-
um nitrate by E. spinifera. Results on the other
biochemical reactions confirm those reported
NOTES 977
Physiological activities
Gelatin Paraffin Na2NO3 Maximum
lique- utiliza- utiliza- growth
tion temp (°C)
- - - - + 42
- - - - - + 42
- - - - - + 38
- - - - - + 38b
- - - - - - 42
- - - - - - 42
earlier by various investigators (5, 11, 17-20).
Except for the maximum growth temperature of
E. spinifera, which we found to be lower than
that reported by Conti-Diaz et al. (5) (38 as
against 41°C), the temperature results were con-
sistent with the published data (5, 11, 17-19).
The ability to hydrolize casein and the inability
to decompose tyrosine are biochemical charac-
teristics that differentiate E. werneckii from E.
jeanselmei, E. spinifera, and W. dermatitidis.
They are additional supportive evidence of dif-
ferences among these species.
Results of the morphological and biochemical
studies show that the isolates of E. werneckii
evaluated in this study, from human and envi-
ronmental origins, are identical to typical iso-
lates recovered from patients with tinea nigra.
The diverse origins and halophilic nature of our
isolates indicate the ubiquitous nature of E.
werneckii.
I thank Libero Ajello for furnishing the tinea nigra isolates
and Ismael A. Conti-Diaz for the IHM isolates, and appreciate
the technical assistance of M. S. Barreto da Silva and
R. C. C. Luizao.
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