Annals of RSCB Vol.

XVII, Issue 1/2012

MORPHOLOGY AND ULTRASTRUCTURE ASPECTS IN SPECIES

BELONGS TO TRICHOPHYTON GENUS USING LIGHT AND
SCANNING ELECTRON MICROSCOPY
C. V. Mihali1, A. Buruiana2, Violeta Turcus1,2, Aurelia Covaci1,
A. Ardelean1,2
1 – INSTITUTE OF LIFE SCIENCES, UVVG ARAD
2 – THE FACULTY OF NATURAL SCIENCES, ENGINEERING AND INFORMATICS,
UVVG ARAD

Summary
The present paper presents the morphology features of the macroconidia, microconidia,
thalus and general aspect of mycelium in Trichophyton genus. The species studied were
Trichophyton mentagrophytes ssp. mentagrophytes and Trichophyton mentagrophytes
ssp. quinckeanum. These species belongs to Arthrodermataceae family, order
Onygenales of Ascomycota division from Fungi. Observations were made on fungal
fragments from culture media containing strains isolated from patients with clinical
suspicious of tinea corporis and tinea capitis. Trichophyton species are causative agents
of dermatophytosis, onychomycosis, tinea barbae, tinea capitis, tinea corporis, tinea
cruris, tinea faciei, and tinea pedis. In species belonging to Trichophyton genus, the
morphological aspects can be analysed using an scanning electron microscope with
environmental mode of observation (ESEM) for ultrastructural details in
micro/macroconidia and talus aspects and general overview images using light
microscopy.
Keywords: ESEM, ultrastructure, light microscopy, morphology, Trichophyton genus.

mihaliciprian@yahoo.com

Introduction tissues, it is considered as one of the main

Trichophyton is a dermatophyte fungus
which can be isolated from the soil,
humans, or animals. The Trichophyton
genus includes anthropophilic, geophilic,
and zoophilic species (De George et al.,
2001; Georg, 1959). Some of the
Trichophyton species are cosmopolitan and
others have a limited geographic
distribution. There are 22 species under
genus Trichophyton. Most common are
Trichophyton mentagrophytes,
Trichophyton concentricum, Trichophyton
rubrum, Trichophyton schoenleinii,
Trichophyton tonsurans, Trichophyton
verrucosum, and Trichophyton violaceum.
From the total number of species, 11 are
often associated with tinea of the scalp, the
nails, and the skin in humans and 4 are
often isolated from animals. Trichophyton
genus is a keratinophilic filamentous fungus
which has the ability to invade keratinized
causes of hair, skin, and nail infections in
humans
(Weitzman et al., 1988). They have several
enzymes, for instance – folic acid,
proteinases, elastase, keratinases. Other
enzymes such as proteinases are the major
virulence factors of Trichophyton species.
Trichophyton mentagrophytes a zoophilic
dermatophyte has 5 five different ssp., T.
mentagrophytes ssp. interdigitale
anthropophilic, T. mentagrophytes var
nodulare a rare anthropophilic form
isolated from cases of tinea pedis, T.
mentagrophytes ssp. mentagrophytes, T.
mentagrophytes ssp. quinckeanum and T.
mentagrophytes var erinacei are zoophilic
dermatophytes. All these ssp. shaping
together the T. mentagrophytes complex.
Differentiation between these ssp. not be
achieved on a single culture medium

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Annals of RSCB
(Ajello and Cheng, 1967; Houck et al.
1996).
The Trichophyton species have
teleomorphic forms and these teleomorphs
are classified under genus Arthroderma,
two perfect states have been described for
T. mentagrophytes: Arthroderma
benhamiae and Arthroderma
vanbreuseghemii (Ajello and Cheng, 1967;
Takashio, 1973). Takashio (Takashio,
1972) divided A. benhamiae in 2 ssp., an
American-European ssp. and an African
ssp. He observed that intra-ssp. matings
were very fertile but inter-ssp. matings were
only low or poorly fertile or sterile. He
observed that the ssp. erinacei which
belongs to the African ssp. and ssp. caviae
which belongs to the American-European
ssp. were sexually degenerated by
comparing with the normal strain of A.
benhamiae. The Finnish and Swedish
strains of T. mentagrophytes were
compatible with A. benhamiae (+ type) and
Material and methods
Were examinated 2 strains of
Trichophyton mentagrophytes ssp.
mentagrophytes and 1 of Trichophyton
mentagrophytes ssp. quinckeanum from
patients with isolated cutaneous lesions of
tinea corporis and tinea capitis, pedis,
barbae. The fragments of fungal
dermatophytes culture media containing
strains isolated from patients with clinical
suspicious of tinea corporis and tinea
capitis, barbae were examinated using
optical microscopy and (ESEM)
environmental scanning electron
microscopy (Ludwig, 1998; Patrick, 2009),
for morphological and structural
particularities studies (Glauert, 1974).
Light microscopy - native samples was
made using adhesive-tape method of
sampling, the morphology studies was
made using a BX 43 Olympus optical
microscope with semi plan apochromate
objectives 10X, 20X, 100X mages were
captured using a digital CCD camera, CX3
Olympus. The stain method used was with
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Vol. XVII, Issue 1/2012
support the European dominance of A.
benhamiae. Numerous variations of colony
morphology are seen among the
Trichophyton mentagrophytes complex
(Beneke and Rogers, 1971; Rippon, 1982).
Macroscopic appearance of the colony -
growth rate may range from being slow to
moderately rapid, colonies are waxy,
glabrous, downy to cottony. The surface
colony color ranges from white to bright
yellowish beige or red violet and the
reverse may be pale, yellowish, brown, or
red – brown. Macroconidia are multicellular
(with 2 or more cells), in some cases can be
smooth, thin or thick walled, cylindrical,
club shaped, or cigar shaped, and are often
absent. Microconidia are unicellular, round
to pyriform shaped, numerously present,
may be solitary or arranged in grape like
clusters.
one drop of lactophenol cotton blue
between the glas slide and adhesive-tape.
ESEM with cooling stage - colony sample
of Microsporum gypseum and
Microsporum canis were excised and
placed in Eppendorf tubes with
Glutaraldehyde 2,7% for 6 hr at 4ºC. After
they were mounted into the Cooling stage
(which is a temperature controlled
specimen holder of ESEM microscope)
which has 3°C and 100% RH . The
morphology studies was made using Fei
Quanta 250 ESEM, the parameters of
ESEM examination were represented by
the following items: the image mode, the
working distance (WD), the beam
conditions, chamber pressure and the
relative humidity into sample. In the same
order they were: the gaseous secondary
electron detector detector (GSED); the
working distance: 5 mm; the beam
conditions: 12,5 – 15 kV, spot 4,5; the
pressure range: 910 to 1400 Pa and the
relative humidity of 90 - 100%.
Annals of RSCB
Results and discussions
Trichophyton mentagrophytes ssp.
mentagrophytes showed a flat rapidly
growing granular whyte colony. Mycelium
was sparse and the powdery or granular
appearance is due to the production of
numerous microconidia. Numerous
variations of shape colony morphology
were seen among the Trichophyton
mentagrophytes complex (Fig. 1 A-C).
Macroscopic appearance of the colony
colours ranges from white to bright
yellowish beige. The colony colours were
Fig.1 – T. mentagrophytes ssp. mentagrophytes, macroscopic appearance of the colony (A,C -
reverse; B,D – obverse), colours ranges from white to bright yellowish beige.
mentagrophytes ssp. mentagrophytes, many
microconidia and macroconidia (Fig.2). In
Trichophyton mentagrophytes ssp.
quinckeanum, the presence of numerous
microconidia and macroconidia were
observed (Fig.3) The numbers of
macroconidia in T.m. ssp. mentagrophytes
were lower compared with T.m. ssp.
quinckeanum. In examined sample from
T.m. ssp. quinckeanum were not met spiral
hyphae. Macroconidia in T.m. ssp.
mentagrophytes are multilocular with
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Vol. XVII, Issue 1/2012
white on obverse side and beige with a
central brown spot on reverse side.
In Trichophyton mentagrophytes ssp.
quinckeanum, the colony was well defined,
The colony has a white higher central area
while the active growing area is superficial
at the surface of the medium. In the
examination with light microscopy, we
observed the following morphology
aspects: a general overview showed the
presence of numerous spiral hyphae in
Trichophyton
smooth walls. In some macroconidia the
apex were pointed tip and others with
rounded terminal cell. Large and elongate
macroconidia were seen. The microconidia
were globular. Macroconidia in T.m. ssp.
quinckeanum, were not so elongate as in
anterior ssp. and they have multilocular
chambers with smooth walls. The branched
thalus has terminal microconidia,
elongated/pyriform in shape. Microconidia
number is smaller than in the T.m. ssp.
mentagrophytes.
Annals of RSCB Vol. XVII, Issue 1/2012

Fig. 2 – T. mentagrophytes ssp. mentagrophytes, macroconidia (A-100X), spiral hiphae (B- 40X),
spiral hiphae / macroconidia – general overview (C – 10X), microconidia/macroconidia (D – 100X).

Fig. 3 - T. mentagrophytes ssp. quinckeanum, macroscopic appearance of the colony (A - reverse; B –

obverse); macro/microconidia (C-100X).

In ESEM analysis could notice the mentagrophytes . In T. mentagrophytes ssp.

difference in colony density between the
analyzed subspecies, the T. mentagrophytes
ssp. mentagrophytes colony has a uniform
appearance with a dense thalus, without
spaces while the T. mentagrophytes ssp.
quinckeanum present discontinuity in the
colony with a thalus multi-branched which
has many cordage (Fig. 4 A-D). In ESEM
investigations weren’t observed
macroconidia. The thalus is without
branches with microconidia arranged in
clusters in T. mentagrophytes ssp.
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quinckeanum thalus is highly branched,
branches are opening wide to thalus (Fig. 4
B-E). In T. mentagrophytes ssp.
quinckeanum the were observed spiral
hyphae, also (Fig. 5).
Microconidia in T. mentagrophytes ssp.
mentagrophytes were arranged in grape
clusters, they were globular in shape while
in the sp T. mentagrophytes ssp.
quinckeanum were elongated/pyriform and
arranged in terminal ramifications.
Annals of RSCB Vol. XVII, Issue 1/2012

Fig. 4 – T. mentagrophytes ssp. mentagrophytes (A- mycelium general aspect; B – thalus with
microconidia; C- microconidia). T. mentagrophytes ssp. quinckeanum (D- mycelium general aspect;
E- branched thalus with terminal microconidia; F- microconidia detail).

Fig. 5 - T. mentagrophytes ssp. quinckeanum, thalus, spiral hiphae, microconidia.

Conclusions
By comparing the two ssp., they have In T. mentagrophytes ssp. mentagrophytes
the following differences: T. species were a greater number of
mentagrophytes ssp. mentagrophytes microconidia, spiral hyphae were observed
presents a less branched thalus with in greater numbers in T. mentagrophytes
abundant microconidia arranged in a cluster ssp. quinckeanum in ESEM investigations .
along the entire tall while in T. Colony aspect in T. mentagrophytes ssp.
mentagrophytes ssp. quinckeanum thalus is quinckeanum presents cordage and small
highly branched, branches are opening wide spaces between the mycelium hyphae while
to thalus. Microconidia are arranged on the in T. mentagrophytes ssp. mentagrophytes
terminal branches of thalus. the colony has a dense uniform appearance.
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Annals of RSCB
Acknowledgements
This work was cofinanced from the
European Social Fund through Sectoral
Operational Programme Human Resources
Development 2007-2013, project number
POSDRU/89/1.5/S/63258 ”Postdoctoral school
for zootechnical biodiversity and food
biotehnology based on the eco-economy and the
bio-economy required by eco-san-genesys”.
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