Sei sulla pagina 1di 24

THE ORDER

ONYGENALES
Introduction

The Onygenales is an order of Ascomycota encompassing species with gymnothecial or


cleistothecial ascomata, evanescent asci, unicellular ascospores and aleurio- or arthroconidial
anamorphs. As circumscribed by Currah (1985). This Order includes four families:
Arthrodermataceae, Onygenaceae, Gymnoascaceae, and Myxotrichaceae.

The family Arthrodermataceae consists of the infamous dermatophytes which have


keratinophilic nature and are responsible for one of the most common human fungal infectious
diseases in the world known as dermatophytoses. The members of family Onygenaceae have
great ecological importance because of their ability to breakdown keratin. The important
systemic dimorphic pathogens are also members of this family. The Gymnoascaceae is a
heterogeneous assemblage of taxa with unknown or poorly defined substrate preferences, and
usually inhabit soil rich in organic matter. Several species of gymnoascaceous fungi had also
found to be able to degrade keratin. The Myxotrichaceae differs from the other onygenalean
families because it is cellulolytic rather than keratinolytic. This family was placed in the
Onygenales (Currah, 1985) but now recognized to be better disposed among the inoperculate
discomycetes (Currah, 1997 and Sugiyama et al., 1999).

There are many reasons for the interest in examination of Onygenalean fungi in the
environment including their clinical interest, hygienic and epidemiological importance as well as
their biotechnological and industrial applications. The order Onygenales is important from the
medical perspective because it includes the true fungal pathogens of humans and animals (i.e.,
the dermatophytes and the dimorphic fungi capable of causing disease in a healthy host). In
addition to the true pathogenic fungi, there are many other onygenalean fungi which survive
saprophytically on soil and wild animals including birds, providing inoculums for primary and
secondary infections to various animals and human. Studies of these fungi in the environment are
therefore, of hygienic and epidemiological importance (Garg et al., 1985 and Sigler, 2002).

Keratinolytic Onygenalean fungi play an important role in the decomposition of the


substrata in the habitats and could be used in many biotechnological applications, for example
they might be used as bioindicators of environmental waste pollution (Ulfig, 2000), tools for

1
bioremediation of keratinic wastes (Singh, 2002), sources of extracellular keratinase enzymes
(Anbu et al,. 2008), indicators of hydrocarbon contamination and bioremediation progress (Ulfig
et al., 2003), and as tools for biodegradation of petroleum hydrocarbon (Ulfig et al., 2006).

Phylogeny and Identification of Onygenalean fungi primarily requires the study of


ascomatal structure, peridial hyphae and appendages, ascospore shape, size and ornamentation,
anamorphic stages and substrate preferences in addition to physiological characteristics (Currah
1985, 1994). However, the members of this order can not always be separated employing these
features and consequently placement of species within the Onygenales can also be problematic in
the absence of a significant distinguishing characters.

Due to the difficulties of the classical (morphological) methods in the distinction of taxa in
this poorly-differentiated fungal group, molecular techniques have been used as a complementary
tool to confirm their identification.

Recently, molecular approaches, mainly based on analysis of different ribosomal DNA


gene regions, have been introduced for the systematic of Onygenales and have proved to be a
reliable tool for clarifying the phylogenetic relationships among this order, especially in
suprageneric relationships (Bowman and Taylor, 1993).

I. Characteristics of the order Onygenales:

According to Currah (1985 & 1994) and Sigler (2002) the Onygenales is an order of
Ascomycota, belonging to the class Eurotiomycetes, which is characterized by formation of
prototunicate asci with eight ascospores and by ascospores that are small (usually less than 8 m
in length), single-celled, and light colored (never dark brown or black). Prototunicate asci are
usually globose or subglobose; they are irregularly arranged within the ascomata and their cell
walls lyse at or near maturity (evanescent walls), allowing for passive discharge of the
ascospores. Ascomata have varied morphologies ranging from nonostiolate, usually globose,
pseudoparenchymatous structures (i.e., cleistothecia) to a loose network (mesh-like) of
differentiated hyphal cells or of branched hyphae (i.e., often called gymnothecia) that sometimes
extend into elaborate hooked, spiralled, or branched appendages. Anamorphs are solitary, single-,

2
or multicelled aleurioconidia or alternate arthroconidia. Several taxa degrade keratin, some are
important human pathogens, and others can degrade cellulose.

This Order has traditionally been divided into four families: Arthrodermataceae,
Onygenaceae, Gymnoascaceae, and Myxotrichaceae. Characters that have been used to separate
families include features of the ascospore wall ornamentation, correlated to some extent with
ascospore shape, habitat, capacity to degrade keratinous or cellulosic substrates or lacking these
enzymatic capacities, and features of the anamorph.

Family Arthrodermataceae

Members of the Arthrodermataceae have keratinophilic nature and are often pathogenic
for human and animals. Ascospore walls are smooth. Anamorphs are aleurioconidia that are
multicelled and /or single-celled and placed in the genera Epidermophyton, Microsporum and
Trichophyton.

Family Onygenaceae

Members of the Onygenaceae have an ability to degrade keratinaceous substrates.


Ascospore walls are ornamented with small pits or depressions (also called puncta), with netlike
ridges (reticulate) or a combination of both, or with minute spiny projections (muriculate).
Anamorphs are aleurioconidia that are predominantly single-celled and placed in the genera
Histoplasma, Blastomyces, Emmonsia, Paracoccidioides, and Chrysosporium, or are alternate
arthroconidia and placed in the genus Malbranchea.

Family Gymnoascaceae

Members of the Gymnoascaceae are a diverse group. They are not keratinolytic and not or
only weakly cellulolytic as determined by in vitro methods. Ascospores are smooth or
ornamented with thickened knobs and are oblate or discoid, sometimes with one or two
longitudinal ridges. Anamorphs are lacking for many species or are arthroconidia or
aleurioconidia.

3
Family Myxotrichaceae

Members of the Myxotrichaceae have fusiform or ellipsoidal ascospores that are striate or
smooth; they degrade cellulosic substrates, and some have Geomyces or Malbranchea
anamorphs. This family was described by Currah (1985) within the Onygenales. Later, Currah
(1997) himself pointed out that Myxotrichaceae is dissimilar to other onygenalean families
because of their affinity to plant debris and cellulolytic characters. This hypothesis was supported
by recent molecular studies that place members of this family closer to the inoperculate
discomycetes. However, the family Myxotrichaceae are included within most reports taking
about the order Onygenales just for convenience (Sigler, 2002)

II. Ecology of onygenalean fungi

Family Arthrodermataceae consists of the genera Arthroderma (anamorphs in


Trichophyton), Nannizzia (anamorphs in Microsporum) and an additional member of the group
known only in its conidial state, Epidermophyton, the infamous dermatophytes which have
keratinophilic nature and are responsible for one of the most common human fungal infectious
diseases in the world (dermatophytoses). The dermatophytes have been divided into three
ecological groups: geophiles, zoophiles and anthropophiles. Probably some of these fungal
pathogens in evolving from their natural habitat in the soil have developed host specificity,
resulting in these three groups. Geophiles are primarily soil-inhabiting and only rarely
encountered as agents of ringworm, with the exception of M. gypseum. Zoophiles are essentially
animal pathogens, although they may cause infection in humans. Anthropophiles are restricted to
man, very rarely infecting animals (Rippon, 1982).

Family Onygenaceae contains an ecologically important group of fungi (keratinophilic


fungi) that can cycle keratin, which is one of the most abundant and highly stable animal proteins
on the earth. Keratin occurs in the form of a wide variety of substances performing many

4
different functions: the claws and armour of reptiles, the feather and beaks of birds, and the
hooves, horns, skin, wool, hair and nails of mammals (Sharma. and Rajak, 2003).

keratinophilic fungi usually inhabit soil of a variety of sites frequented by animals and man
such as playgrounds, recreation centers, lake side, beaches, poultry farms, breeding farms,
zoological parks, animal cages, forest and cultivated fields. (Garg et al., 1985, Sundaram, 1987,
Ali-Shtayeh , 1989, Shukia et al., 2003, Moallaei et al., 2006, Irum, et al., 2007 &
Mahmoudabadi1and Zarrin, 2008) Besides soil, birds' feathers and nests (Dixit and Kushwaha,
1991 & Efuntoye, 2001), hairs of wild animals (Ali-Shtayeh et al., 1988), air dust (Vidyasagar et
al., 2005), water (Ali-Shtayeh et al., 2002), plant debris (Currah, 1985), sewage (Ulfig et al.,
1996) and dung (Currah, 1985) are other habitats for the survival of keratinophilic fungi.

The important dimorphic pathogenic fungi which cause systemic mycoses, Histoplasma
capsulatum, Blastomyces dermatitidis, Coccidioides immitis and Paracoccidioides brasiliensis
have been classified in the Onygenaceae family. None of them display keratinolytic activity
which is a mycological phenotype of many fungal species within Onygenales.

Histoplasma capsulatum, is endemic to the temperate zones of the world, including the
Americas, Asia, and Africa (Quinet-Leimann et al., 2005). It is highly endemic to the Ohio
and Mississippi river valleys of the United States. Histoplasma capsulatum, grows in soil
associated with the accumulations of bird and bat droppings (Cano and Hajjeh, 2001).

The disease caused by Blastomyces dermatitidis occurs primarily in the southeastern and
south-central states of U.S.A. bordering on the Mississippi and Ohio rivers basins (Klein et al.,
1986,a). Worldwide cases have been reported (Schutze et al., 1998). However, the great majority
of cases are from the eastern portion of North America. B. dermatitidis is nourished on acid soils
with high organic content, abundant moisture, and possibly enrichment by animal droppings
(Klein et al., 1986,b & Klein et al., 1987).

The endemic region for Coccidioides immitis lies exclusively in the Western Hemisphere
(Hector et al., 2005). This life zone corresponds with the hot deserts of the southwestern United
States and northwestern Mexico. Cases of coccidioidomycosis may also arise outside endemic
areas, related to a recent visit to an endemic area or infection through exposure to fomites from

5
such an area (Desai et al., 2001). Coccidioides is a soil-dwelling organism which prefers alkaline
soil, containing increased nitrogenous waste, in relatively warm dry climates. (Galgiani, 1993
& Pappagianis, 1988).

The geographical distribution of Paracoccidioides brasiliensis is confined to the region


between the tropics (i.e., restricted to Latin America from Mexico to Argentina. Its ecological
niche is still unknown. The fungus has been rarely isolated from the soil and armadillos
(Dasypus novemcinctus) were shown to harbor the fungus in their internal organs (Conti-Diaz
et al., 2007).

Members of Gymnoascaceae do not exhibit strong substrate preferences, and usually


inhabit soil rich in organic matter and decaying vegetations including dead grass, nests, dung,
ect., (Currah, 1985). Several species of gymnoascaceous fungi, Ctenomyces serratus,
Gymnoascus reesii, G. intermedius and Gymnascella dankaliensis are often recovered from soil
by hair-baiting technique. These species can degrade keratin. C. serratus (anamorph
Myceliophthora vellerea) is found on feathers and in soil- harbouring feathers or other
keratinaceous materials (Pugh et al., 1962 & Caretta et al.,1992). G. reessii and G. intermedius
are coprophilous but they are frequently isolated from soil enriched with the dung of herbivores
by hair-baiting technique (de Hoog & Guarro, 1995).

Myxotrichaceae, members of this family can degrade cellulosic substrates and some
species are also known to form mycorrhizas with ericaceous host plants (Currah, 1985).

III. Isolation of onygenalean fungi

Studies of the Onygenales were restricted to the isolation of representatives of the


Arthrodermataceae and Onygenaceae either from clinical samples or from environmental sources
including soil, Keratinous substrates, and Keratin-enriched debris. The most common methods
employed to isolate these fungi from non-clinical sources are baiting methods that use native
keratin, such as hair and feathers (Currah, 1985; Sundaram, 1987; Ali-Shtayeh , 1989; Dixit and
Kushwaha, 1991; Ulfig et al., 1996; Efuntoye, 2001; Shukia et al ., 2003; Moallaei et al.; 2006;
Irum, et al., 2007 & Mahmoudabadi and Zarrin, 2008). Most other taxa in the Onygenales are
only rarely reported because of their limited competitive activity which seriously hampered their

6
isolation by ordinary isolation techniques. Consequently, little is known about their natural
habitat or the frequency of their occurrence in nature (Kuehn et al., 1961; Orr et al., 1977;
Guarro et al., 1992; Uchiyama et al., 1995; Udagawa, and Uchiyama, 1999 and Udagawa and
Uchiyama, 2000 a, b).

Methods of isolation:

A. Dermatophytes:

1. From patients: Scrapings of infected skin, hair and infected nail cuttings were collected using a
sterilized scalpel or nail clipper. These samples are cultured on plates of Sabouraud dextrose agar
containing chloramphenicol and cycloheximide. Plates were incubated at 2528 C for 4-6 weeks
(Weitzman et al., 1988).
2. From infected animals: Skin scales were collected by scraping of the margin of the
lesion using a sterile scalpel blade into sterile Petri dish. Hairs were collected by removing dull
broken hairs from the margin of the lesion using sterile tweezers as described (Cheesbrough,
1992). Each sample collected is cultured on of Sabouraud dextrose agar containing
chloramphenicol and cycloheximide, incubated at 28 C for 4-6 weeks.
3. From soil: via hair bating technique

B. Keratinophilic fungi:

Collection of samples: samples can be collected from any substrate enriched in keratin
including:

1. Soil: sites frequent by animals and humans like poultry farms, breeding farms, zoological
parks, public parks, schools, play grounds, marketplaces, etc.
2. Bird feathers
3. Animal hairs and claws
4. Herbivore and carnivore dung
5. Sewage
6. Wastes
7. house dust

7
Isolation procedure:

1. From soil (including wastes, sewage sediments, dung): The keratinolytic nature of these
fungi makes it possible to isolate them from soil by implanting hair, the hair baiting
technique initially developed by Vanbreuseghem (1952). Isolation of keratinophilic fungi
can also be done by the other techniques such as the dilution plate method (also used for
isolation from floor dust) or soil plate method although the hair baiting method is better.
2. From animal hair and claws and bird feathers: These samples were sprinkled over the
surface of plates half filled with double sterilized soil. The soil was moistened with
distilled water. The plates were incubated at 25 C for 4-6 weeks. Samples of hairs, claws
and feathers showing fungal growth were transferred to surface of on plates of Sabourauds
dextrose agar sublemented with chloramphenicol and cycloheximide, incubated at 25C
till complete fungal growth.

C. Dimorphic fungi:

1. From patients: Sputum or other respiratory secretions are taken from patient having
pulmonary infection. For patients who have disseminated infection, samples taken from
blood, bone marrow, liver tissue, CNF, skin or mucosal lesions. These samples are
cultured on a variety of artificial media, including blood agar. Plates were incubated at 30
C for 4-6 weeks. The growth of dimorphic fungi on artificial media represents a
laboratory hazard and suspected samples should be handled accordingly (Warnock, 2000).
2. From soil: The procedure of Emmons (1942) was employed in isolating the systemic
pathogenic fungi from the soil by injecting the supernatant from soil suspensions
intraperitoneally into mice or guinea pigs. In addition to the mouse inoculation (in vivo),
there are various in vitro methods used for isolation of the dimorphic fungus Coccidioides
immitis from soil.

Mouse Inoculation Procedure: After thorough mixing, 1 heaping teaspoonful of soil was
placed in 30 ml of physiological saline that contained 5,000 units of penicillin G and 1,000
units of streptomycin per ml. After vigorous stirring, the soil was allowed to settle for
approximately one hour. Five ml of supernatant were then pipetted off and 1 ml injected
intraperitonealty into each of 4 mice. Eight weeks after inoculation, the mice were

8
sacrificed and small portions of their liver and spleen, plus any observable abscesses or
lesions, were cultured on plates of Sabouraud's dextrose agar and brain heart infusion
blood agar. The plates were incubated at 25 C and examined periodically for the
development of pathogenic fungi.

In vitro methods for isolation of Coccidioides immitis from soil:

1. Spray method of Swatek (1956): Five ml of soil suspension (1: 100) was sprayed onto
the surface of five plates of Sabouraud's Agar, using alcohol sterilized "Windex"
sprayers. 9 ml of sterile water (1: 100) and mixed by inversion. One ml of each 1: 100
dilution of a suspension was added to a tube of sterile water (1: 1000) and mixed by
inversion. Five ml of each of the 1: 100 and 1: 1000 dilutions of suspensions were
sprayed onto five plates of medium in the same manner as were the 1: 10 aliquots. In an
upright position the plates were incubated for 24 hours at 37C to dry and then for two
weeks at 30 C.
2. Flotation-plate method of Walch et al. (1961): Five ml of soil suspension were mixed
with 5 ml of sterile 0.08 % CuSO 4 solution in a test tube. The mixture was incubated
at 37 C for 24 hours. One ml this mixture was pipetted onto and spread over the
surface of plates of Walch's Agar medium. the plates were incubated for 24 hours at
37C to dry and then for two weeks at room temperature.
3. Double pour-plate method of Omieczynski et al., (1965): Five ml of soil suspension
was pipetted into ten sterile Petri dishes. Approximately 15 ml of Yeast Extract Agar
was poured into each plate, swirled to mix, and allowed to solidify. A second 15 ml
aliquot of medium was poured into each plate. The plates were incubated at 30C for
two weeks.

D. Cellulolytic fungi:

Collection of samples: samples can be collected from numerous plant debris including woods,
barks and leaves of trees especially in forests.

Isolation procedure: Bait blocks (made of decayed woody debris) were surface sterilized by
briefly flaming, and then spread onto cornmeal agar and malt extract agar. Plates were incubated

9
at room temperature (21C). Hyphae growing from the wood fragments were sub-cultured on
cornmeal agar and oatmeal agar (Sigler et al., 2000).

IV. Identification

Identification of Onygenalean fungi primarily requires the study of ascomatal structure,


peridial hyphae and appendages, ascospore shape, size and ornamentation anamorphic stages and
substrate preferences (Currah, 1985). However, members of this order can not always be easily
separated employing these features. The major difficulty in identification of these fungi comes
when a fungus exists in two stages (sexual and asexual) as both the stages can be
morphologically quite dissimilar. It is very difficult to assign proper genera or species without the
knowledge of both the stages. It becomes more difficult in fungi that are heterothallic [i.e. they
have different mating types (+ and strains) and sexual stage (which results in ascospores)], as
they form sexual stages only when opposite mating types cross.

Heterothallic compatibility is common among onygenalean fungi and the teleomorph can be
obtained only through mating trials. These tests are of great value in confirming identity and in
establishing biological relationships, but are rarely employed because of the need for a library of
appropriate test isolates, and the length of time required for development and maturity of the
sexual structures (Sigler, 2002). For these reasons, the Q-10(H2) ubiquinone system is recently
used for the identification of onygenalean fungi, and also the DNA based identification
techniques are found to be especially useful for onygenalean fungi which are difficult to
distinguish morphologically (Sharma and Rajak, 2003).

Molecular identification:

Due to the difficulties of the classical (morphological) methods in the distinction of taxa
in this poorly-differentiated fungal group, molecular techniques have been used as a
complementary tool to confirm their identification. Recently, several molecular markers have
been used successfully in identification and taxonomy of those fungi. The RFLPs of the
mitochondrial DNA (mtDNA) has been used to corroborate the separation of the different species
of Keratinomyces (Trichophyton) (Guillamon et al. 1996), and to complete morphological criteria
in the identification of several problematic strains of Chrysosporium Corda (Cano et al. 1996).

10
On the other hand, PCR primers based on the internal transcribed spacers (ITS) have been used
by Greene (2000) to identify Coccidioides immitis isolated from soil.

The analysis of specific DNA sequences of the (ITS) also has proven to be useful in
identification of onygenalean fungi and has been used by many investigators to identify several
keratinophilic species (Zaki et al. 2005; Deshmukh and Verekar, 2006 & Deshmukh et al. 2008),
dermatophytes (Mochizuki et al., 1999; Woldeamanuel et al., 2005 and Li et al., 2008) and also
to identify species of Chrysosporium including C. undulatum (Vidal et al. 1999) and C. zonatum
(Abdel-Razik and Zaki, 2008).

V. Phylogeny:

In classifying of the Onygenales, ascoma structure was traditionally emphasized to


circumscribe the taxa (Benjamin, 1956; Kuehn, 1958, 1959; Apinis, 1964; Benny and
Kimbrough, 1980). Currah (1985) revised this order based on a large number of collections and
pointed out that the resemblance of peridial structure does not reflect the phylogenetic lineage
because many of them might result from convergent evolution. He introduced physiological
characteristics in addition to morphological differences of ascospore and conidia for the higher
classification in the Onygenales, and established four families in this order, viz.,
Arthrodermataceae, Gymnoascaceae, Myxotrichaceae, and Onygenaceae. His system has been
widely accepted among mycologists, though von Arx (1987) proposed another system based
mainly on the morphology of ascospores. Von Arx placed the onygenalean taxa into the
Eurotiales, dividing the order among four families, i.e., the Amauroascaceae, Eurotiaceae,
Gymnoascaceae and Onygenaceae.

After Currah's monograph, new taxa or new combinations have been proposed, and
physiological, ecological and molecular data related to the order have been accumulated.

Molecular techniques have been introduced for the systematics of Onygenales and have
proved to be a reliable tool for clarifying the phylogenetic relationships among the Onygenales
(Bowman and Taylor, 1993).

11
With the exception of the Myxotrichaceae, a group now recognized to be more closely
allied to the Leotiales (Currah ,1997, Sugiyama et a.,l 1999), Currahs concept of the Onygenales
has been supported by the results of studies of the molecular systematics and phylogentic
analysis of members of this order.

The Gymnoascaceae, a family once thought to represent a heterogeneous assemblage of


taxa with affinities to the Arthrodermataceae and Eurotiales (Currah, 1985 & 1994), forms a
monophyletic group in phylogenies based on the analysis of nuclear ribosomal RNA (rRNA)
gene sequences (Sugiyama et al., 1999, Sugiyama and Mikawa , 2001, Untereiner et al., 2002).
The phylogenetic relationships within the Gymnoascaceae is still unclear so further sequencing
studies including many gymnoascaceous species is required.

The Arthrodermataceae, which encompasses taxa with smooth ascospores and anamorphs
assigned to Chrysosporium Corda, Epidermophyton Sabour., Microsporum Gruby and
Trichophyton Malmsten, is represented as a well-supported lineage in analyses of nuclear rRNA
and chitin synthase gene sequences (Leclerc et al., 1994, Sugiyama et al .1999 & Herr et al.
2001).

The phylogenetic structure of the Onygenaceae is resolved less clearly. Recent sequence-
based phylogenies indicate that the family is polyphyletic (Sugiyama et al., 1999, Herr et al.,
2001, Sugiyama and Mikawa 2001, Gibas et al., 2002 & Untereiner et al., 2002).

The polyphyletic Onygenaceae was suggested to be composed of three clades; 1) most


members of Onygenaceae, 2) Spiromastix warcupii, and 3) pathogenic dimorphic fungi
(Sugiyama et al., 1999).

One clade recognized consistently in molecular phylogenetic studies of the Onygenaceae


includes a group of medically important taxa encompassing the dimorphic systemic pathogens.
Taxa identified as members of this clade in phylogenies inferred from nuclear small subunit
(SSU) rRNA, nuclear large subunit (LSU) rRNA and internal transcribed spacer (ITS) sequences
include Ajellomyces capsulatus (anamorph Histoplasma capsulatum Darling), A. crescens
(anamorph Emmonsia crescens), A. dermatitidis (anamorph Blastomyces dermatitidis Gilchrist &

12
Stokes) and species of the anamorph genera Emmonsia and Paracoccidioides (Peterson and
Sigler 1998, Sugiyama et al., 1999, Vidal et al., 2000 & Herr et al., 2001).

Untereiner et al. (2004) examined the Phylogenetic structure of the Onygenaceae based on
the analysis of nuclear LSU and ITS rDNA sequences for an expanded set of taxa. Their results
provide further evidence for the recognition of the clade encompassing Ajellomyces (including
the anamorph genera Blastomyces, Emmonsia and Histoplasma) and Paracoccidioides and they
described this clade as a new family known as Ajellomycetaceae.

Spiromastix Kuehn & Orr, a nonpathogenic member of the Onygenaceae, recently was
positioned within this clade based on the comparison of nuclear LSU sequences (Sugiyama and
Mikawa, 2001). This finding was corroborated by Untereiner et al., (2002) in an investigation
that examined phylogenetic relationships of species of Ajellomyces McDonough & Lewis,
Polytolypa Scott & Malloch and Spiromastix inferred from the analysis of nonmolecular
characters and sequences from the nuclear LSU and mitochondrial SSU rRNA genes. Based on
the results of their study, Untereiner et al., (2002) transferred Spiromastix grisea Currah &
Locquin-Linard to Ajellomyces and restricted Spiromastix (typified by S. warcupii) to species
isolated from soil. Polytolypa hystricis, a species described from porcupine dung (Scott et al.,
1993), also was shown to be closely related to Ajellomyces and Spiromastix, but its phylogenetic
position was not sufficiently resolved to propose its transfer to either genus (Untereiner et al.,
2002).

Environmental importance of the order Onygenales:


Onygenalean fungi as human pathogens: Most true pathogenic fungi, with exception
of the black yeasts, belong to this order. The members of Arthrodermataceae, dermatophytes, are
group of morphologically and physiologically related keratinophilic fungi that have the capacity
to invade keratinized tissue (skin, hair & nail) of human and other animals causing
dermatophytoses which commonly referred to as ringworm or tinea.

In the family Onygenaceae, there are numerous pathogenic species, for example,
Aphanoascus spp. which are occasionally agents of superficial infections (Cano and Guarro,
1990) and whose anamorphs are Chrysosporium spp., and Neoarachnotheca keratinophila,

13
which has recently been described as the teleomorph of Myriodontium keratinophilum (Cano et
al., 1997) and which has been reported as causing human sinusitis (Maran et al., 1985).

Other, more pathogenic fungi are the systemic dimorphic pathogens, Blastomyces
dermatitidis, Histoplasma capsulatum, Coccidioides immitis and Paracoccidioides. C. immitis is
the agent that causes coccidioidomycosis, disease start from lung in the form of
bronchopneumonia and can disseminate to other body parts and this disseminated form is often
fatal. Histoplasmosis is a very common granulomatous disease that is caused by the inhalation of
conidia of H. capsulatum and results in a wide range of clinical manifestations as a consequence
of an intracellular infection of the monocyte-macrophage system. P. brasiliensis is the agent of
paracoccidioidomycosis. Initially a primary pulmonary infection that is usually subclinical and it
can progress to a systemic, chronic granulomatous disease. B. dermatitidis is the agent of
blastomycosis, a chonic granulomatous and suppurative disease that may affect lungs, skin, and
mucous membrane, or other organs (Guarro et al., 1999).

In the two remaining families of Onygenales, there are only a few pathogenic species.
Myxotrichum deflexum, a species of doubtful pathogenicity, is found in Myxotrichaceae.
Gymnoascaceae contains Gymnoascus dankaliensis, which has caused some superficial
infections, and Arachnomyces nodosetosus, which was recently proposed to be the teleomorph on
Onychocola canadensis, a pathogenic species found exclusively in human nail and skin lesions
(Sigler et al., 1994).

Hygienic and epidemiological importance of onygenalean fungi: In addition to the true


pathogenic fungi belonging to this order which display potentially pathogenic properties to
animals, including human beings, there are many other onygenalean fungi which survive
saprophytically on soil and wild animals including birds, providing inoculums for primary and
secondary infections to various animals and human. Studies of these fungi in the environment are
therefore, of hygienic and epidemiological importance (Garg et al., 1985).

Keratinolytic onygenealean fungi as bioindicators of environmental waste pollution:


The qualitative and quantitative composition of keratinolytic fungi can be used as a
multifunctional bioindicator of environmental pollution with waste. It means that the

14
composition reflect the complex influence of many environmental factors (Ulfig, 2000). The
most important factors are as follows:

delivery and congregation of keratin remnants and fungal propagules together with the
decomposition of the remnants in a given environment.
level of environmental pollution with human and/or animal faeces.
physico-chemical factors, including climatic conditions.
presence of toxic compounds and elements of industrial and natural origin in the
environment such as detergents, cyanides, phenols, heavy metals, salts, etc.
the infection risk associated with the contamination of the environment with potential
fungal pathogens.

Keratinolytic onygenealean fungi as a tool for bioremediation of keratinic wastes:


Keratin is the structural component of vertebrates skin and its adnexes which is arises as a waste
product in a variety of ways, as a result of shedding and moulting of animal appendages, death of
animals or as a by-product of poultry and leather industry. This keratinic waste is totally wasted
in the waste removal areas or is ploughed into fields. This latter practice is dangerous for the
environment from the public health point of view. It is well known that during uncontrolled
keratin decomposition, especially by anaerobic bacteria, large quantities of toxic substances
including hydrogen sulphide and ammonia is released. There is currently little prospect for clean
up of these wastes by physico-chemical means alone because of the extreme expense, danger and
intensity of labour. In the light of these facts, the problem of searching for alternate methods of
utilization and neutralization of keratinic wastes is very important. In the last few years such
methods as microbiological processing of wastes with high protein content using some
keratinolytic bacteria, actinomycetes and fungi have aroused much interest (Singh, 2002).

Keratinolytic onygenalean fungi as a source of extracellular keratinase enzymes: The


keratinase enzymes have important applications in biotechnological and industrial processes
involving keratin-containing wastes from the poultry and leather industries through the
development of non-polluting processes and dehairing of skin and hides. These waste materials
can also be converted into economically useful feather meal, nitrogenous fertilizers,
biodegradable films, glues, and foils. The production of extracellular keratinase has been

15
reported in various keratinous substrates by many investigators including Anbu et al. (2008) who
purified an extracellular keratinase from Trichophyton sp. HA-2 isolated from feather dumping
soil.

Keratinolytic onygenalean fungi as indicators of hydrocarbon contamination and


bioremediation progress: The inhibitory effect of polar compounds (products of the oil
hydrocarbon biodegradation process) in leachates on the radial growth and dry weight (biomass)
production by keratinolytic fungi especially Trichophyton ajelloi and the qualitative and
quantitative composition of keratinolytic fungal communities can be used as indicators of soil
petroleum hydrocarbon contamination and associated bioremediation progress. (Ulfig et al.,
2003).

Keratinolytic onygenalean fungi as a tool for biodegradation of petroleum


hydrocarbon: keratinolytic fungi are able to remove petroleum hydrocarbon including hexane,
toluene, hexadecane, pristane and autoclaved crude oil from mineral media. These fungi display
lipolytic activity during degradation of protein and the hydrocarbon removal process is
dependent on fungal proteolytic or keratinolytic activity (Ulfig et al., 2006).

References

Abdel-Hafez AI, Moharram AM, Abdel-Gawad KM. (1990, a) Survey of keratinophilic and saprobic
fungi in the cloven-hooves and horns of goats and sheep from Egypt. J Basic Microbiol. 30(1):13-
20.

Abdel-Hafez, S. I., Moubasher, A. H., Barakat, A. (1990, b) Keratinophilic fungi and other moulds
associated with air-dust particles from Egypt. Folia Microbiol (Praha). 35(4):311-25.

Abdel-Hafez, A. I. I.and El-sharouny, H. M. M. (1990) occurrence of keratinophilic fungi in sewage


sludge from Egypt. J. Basic Microbiol. 30, 73.

Abdel-Mallek, A.Y., Bagy, M. M. and Moharram, A. M. (1989) Keratinolytic fungi of wadi Qena in
Egypt. Folia Microbiol (Praha). 34(1):37-41.

Abdel-Razik, M. and Zaki, S. M. (2008) Experimental Pathogenicity and Molecular Characterization


of an Environmental Isolate of Chrysosporium zonatum Al-Musallam and Tan (Family:
Onygenaceae, Order: Onygenales). Int. J. Agri. Biol. 10:(3) 273-7.

16
Abdel-Razik, M., Moawad, F., Abdel-Aziz, M.H. and Shokr, A.E.M. (2001): The spread of
dermatophytes and dermatomycosis between people in the rural areas of Alarish, North Sinai,
Egypt. El-Minia Science, Bulletin. 13(2) & 14(1) 127-138.

Ali-Shtayeh, M. S. (1989) Keratinophilic fungi of school playgrounds in the Nablus area, West Bank of
Jordan. Mycopathologia 106: 103-108.

Ali-Shtayeh, M. S. Arda, H. M. Hassouna, M. and Shaheen S. E. (1988) Keratinophilic fungi on the


hair of cows, donkeys, rabbits, cats, and dogs from the West Bank of Jordan Mycopathologia
104:109-121.

Ali-Shtayeh, M. S., Khaleel, T. Kh. M. and Jamous, R. M. (2002) Ecology of dermatophytes and
other keratinophilic fungi in swimming pools and polluted and unpolluted streams
Mycopathologia 156: 193205.

Anbu, P., Hilda, A., Hwal-Won, S, Byung-Ki, H. and Jayanthi, S. (2008) Extracellular keratinase
from Trichophyton sp. HA-2 isolated from feather dumping soil International Biodeterioration &
Biodegradation xxx 16.

Apinis, A.E. (1964) Revison of British Gymnoascaceae. Mycol. Pap. Commonw. Mycol. Inst. 95: 1-56.

Arx, J. A. v. (1987) A re-evaluation of the Eurotiales. Persoonia 13: 273-300.

Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struthl, K.
(1999) Current Protocols in Molecular Biology. John Wiley and Sons, New York.

Benjamin, R.K. (1956) A new genus of the Gymnoascaceae with a review of the other genera. El Alis o
3: 301-328.

Benny, G. L. and Kimbrough, J.W. (1980) A synopsis of the orders and families of plectomycetes with
keys to genera. Mycotaxon 12: 1-91.

Bowman, B. H. and Taylor, J.W. (1993) Molecular phylogeny of pathogenic and non-pathogenic
Onygenales. In: The fungal holomorph: Mitotic, meiotic and pleomorphic speciation in fungal
systematics, (ed. Reynolds, D. R. and Taylor, J. W.), pp. 169-178. CAB International,
Wallingford.

Cano, J. and Guarro, J. (1990) The genus Aphanoascus. Mycol. Res. 94: 355377.

Cano, J., Guillamn, J. M., Vidal, P. and Guarro, J. (1996) The utility of mitochondrial DNA
restriction analysis in the classification of strains of Chrysosporium (hyphomycetes).
Mycopathologia. 134(2):65-9.

Cano, J., K. Ulfig, J. M. Guillamon, P. Vidal, and J. Guarro (1997) Studies on keratinophilic fungi.
IX. Neoarachnotheca gen. nov. and a new species of Nannizziopsis. Antonie Leeuwenhoek Int. J.
Genet. 72:149158.

Cano, J., Vidal, P., Guarro, J., Castaneda Ruiz, R. F. and de Vroey, C. H. (1996) Three new species
of Amauroascus from tropical regions. Mycological Research 100: 343348.

Cano, M..V..C. and Hajjeh, R. A. (2001) The epidemiology of histoplasmosis: a review. Semin Respir
Dis, 16: 109-118.

17
Caretta, G., Mangiarotti AM and Piontelli E. (1992) Keratinophilic fungi isolated from soil of Italian
parks in the province of Pavia. Eur J Epidemiol 8:330-339.

Cheesbrough, M., (1992) Medical laboratory manual for tropical countries.Volum 2. Tropical health
technology, Butterworth-Heinemann, Great Britain, pp. 371-385.

Conti-Diaz IA. (1989) On the unknown ecological niche of Paracoccidioides brasiliensis. Our
hypothesis of present status and perspectives. Rev Inst Med Trop S Paulo 2007; 49: 131-134.

Currah, R. S. (1994) Peridial morphology and evolution in the prototunicate ascomycetes. In:
Hawksworth DL, ed. Ascomycete systematics: problems and perspectives in the nineties. New
York: Plenum Press. p. 281293.

Currah, R. S. (1997) Taxonomy of saprophytic and pathogenic fungi in the Onygenales. Annual Report
of the Research Center for Pathogenic Fungi and Microbial Toxicosis. Chiba University: Japan. p.
4454.

Currahm R. S. (1985) Taxonomy of the Onygenales: Arthrodermataceae, Gymnoascaceae,


Myxotrichaceae and Onygenaceae. Mycotaxon 14:1216.

de Hoog, G. S. and Guarro, J. (1995) Atlas of Clinical Fungi. Centraalbureau voor Schimelcultures /
Universitat Rovira I Virgili, 134, 235, 278, 290, 388,394, 392, 430, 452, 632, 634, 636, 638.

Desai S. A., Minai, O. A., Gordon, S. M., ONeil, B, Wiedeman H. P. and Arroliga, A. C. (2001)
Coccidioidomycosis in non endemic areas: a case series. Respir Med 95: 305-309.

Deshmukh, S. K. and Verekar, S. A. (2006) Keratinophilic fungi from the vicinity of meteorite crater
soils of Lonar, India. Mycopathologia. 162(4):303-6.

Deshmukh, S. K., Mandeel, Q. A. and Verekar, S. A. (2008) Keratinophilic fungi from selected soils
of Bahrain. Mycopathologia. 165(3):143-7.

Dixit, A.K. and Kushwaha, R.K.S. (1991) Occurrence of Keratinophilic Fungi on Indian Birds. Folia
Microbiol. 36 (4), 383 - 386

Efuntoye, M. O. (2001) Occurrence of keratinophilic fungi and dermatophytes on domestic birds in


Nigeria Mycopathologia 153: 8789.

El-Zayat, S.A. (2000): A study dermatophytosis in Aswan. Az. J. Microbiol.49:119-127.

Emmons, C. W. (1942) Isolation of Coccidioides from soil and rodents. Public Health Rept. 57: 109--
111.

Felsenstein, J. (1981) Evolutionary trees from DNA sequences: a maximum likehood approach. J. Mol.
Evol. 17:368-367.

Furuya, K., and Naito, A. (1980) Stimulation of ascospore germination by phenolic compounds in
members of the Sordariaceae. Transactions of the Mycological Society of Japan 21:7785.

Galgiani, J. N. (1993) Coccidioidomycosis. West J Med 159: 153-171.

Garg, A. P., Gandotra, S., Mukerji, K. G. and Pugh, G. J. F. (1985) Ecology of keratinophilic fungi.
Proc. Indian Acad. Sci. (plant Sci.),94: (2, 3) 149-163.

18
Gibas, C. F.C., Sigler, L., Summerbell, R. C. and Currah, R. S. (2002) Phylogeny of the genus
Arachnomyces and the establishment of Arachnomycetales, a new eurotiomycete order in the
Ascomycota. Stud Mycol 47:131139.

Graser, Y.; El Fari, M., Vilgalys, R., Kuijpers, A. F., De Hoog, G. S., Presber, W. and Tietz, H.
(1999): Phylogeny and taxonomy of the family Arthrodermataceae (dermatophytes) using
sequence analysis of the ribosomal ITS region. Med Mycol. 37(2):105-14.

Greene, D. R. (2000) Soil isolation and molecular identification of Coccidioides immitis. Mycologia 92:
( 3) 406410.

Guarro, J., Summerbell, R.C. and Samson, R.A., eds. (2003) Onygenales: The Dermatophytes,
Dimorphicsand Keratin Degraders in their Evolutionary Context,Studies in Mycology, Vol. 47.
Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands.

Guarro, J., Gen, J. and De Vroey, C. (1992) Amaurascopsis, a new genus of Eurotiales. Mycotaxon
45:171178.

Guarro, J., Gene, J. and Stchigel, A. M. (1999) Developments in Fungal Taxonomy. Clin.Microbiol.
Rev. 12(3):254-500.

Guillamn, J. M., Cano, J., Ramn, D. and Guarro, J. (1996) Molecular differentiation of
Keratinomyces (Trichophyton) species. Antonie Van Leeuwenhoek. 69(3):223-7.

Hector, R. F. and Laniado-Laborin R. (2005) Coccidioidomycosis-A fungal disease of the Americas.


PLoS Med. 2: 2e.

Herr, R. A., Taracha, E. J., Taborda, P. R., Taylor, J. W., Ajello, L. and Mendoza, L. (2001)
Phylogenetic analysis of Lacazia loboi places this previously uncharacterized pathogen within the
dimorphic Onygenales. J Clin Micro 39:309314.

Irum, F., Suhail, M. and Aero, H. (2007) Keratinophilic Fungi from the Soil of District, Jamshoro,
Sindh, Pakistan. Pak. J. Bot., 39(4): 1377-1382.

Jiang, B., Bartlett, M. S., Allen, S. D., et al. (2000) typing of Histoplasma capsulatum isolates based
on nucleotide sequence variationin the internal transcriped spacer regions of rRNA genes. J Clin
Microbiol. 38: 241_245.

Klein, B. S., Vergeront, J. M. and Davis, J. P. (1986, a) Epidemiological aspects of blastomycosis, the
enigmatic systemic mycosis. Semin Respir Infect 1:29-39.

Klein, B. S., Vergeront, J. M. and DiSalvo, A. F., (1987) Two outbreaks of blastomycosis along rivers
in Wisconsin: isolation of Blastomycosis dermatitidis from riverbank soil and evidence of
transmission along waterways. Am Rev Respir Dis 136:1333-8.

Klein, B.S., Vergeront, J. M. and Weeks, R. J. (1986, b) Isolation of Blastomycosis dermatitidis in soil
associated with a large outbreak of blastomycosis in Wisconsin. N Engl J Med 314:529-34.

Kuehn, H. H., Orr, G. F. and Ghosh, G. R. (1961) A new and widely distributed species of.
Pseudoarachniotus. Mycopathologia, 14, 215-229

19
Kuehn, H.H. (1958) A preliminary survey of the Gymnoascaceae. I. Mycologia 50: 417-439. Kuehn,
H.H. 1959. A preliminary survey of the Gymnoascaceae. II. Mycologia 51: 665-692.

Leclerc, M. C., Phillipe, H. and Gueho, E. (1994) Phylogeny of dermatophytes and dimorphic fungi
based on large subunit ribosomal RNA sequence comparisons. J Med Veter Mycol 32:331341.

Li, H. C., Bouchara, J. .,P, Hsu, M. M., Barton, R., Su, S. and Chang, T. C. (2008)
Identification of dermatophytes by sequence analysis of the rRNA gene internal transcribed
spacer regions. J Med Microbiol. 57(5):592-600.

Maghraby, T., A., Gherbawy, T. A., Y. A. M. H. and Hussein, M. A. (2008) Keratinophilic fungi
inhabiting floor dusts of student houses at the South Valley University in Egypt. Aerobiologia
24:99106.

Mahmoud A.L. (1993) Dermatophytes and other associated fungi isolated from ringworm lesions of
camels. Folia Microbiol (Praha). 38(6):505-8.

Mahmoud, A. L. (1995) Dermatophytes and other keratinophilic fungi causing ringworm of horses.
Folia Microbiol (Praha). 40(3):293-6.

Mahmoudabadi1, A. Z. and Zarrin, M. (2008) Isolation of dermatophytes and related keratinophilic


fungi from the two public parks in Ahvaz Jundishapur Journal of Microbiology 1(1): 20-23 20.

Maran, A. G. D., Kwong, K. Milne, L. J. R. and Lamb, D. (1985) Frontal sinusitis caused by
Myriodontium keratinophilum. Br. Med. J. 20:207.

Moallaei , H., Zaini, F., Pihet, M., Mahmoudi, M. and Hashemi, J. (2006) Isolation of Keratinophilic
Fungi from Soil Samples of Forests and Farm Yards Iranian J Publ Health, 35 (4):62-69.

Mochizuki, T., Kawasaki, M., Ishizaki, H. and Makimura, K. (1999) Identification of several clinical
isolates of dermatophytes based on the nucleotide sequence of internal transcribed spacer 1 (ITS
1) in nuclear ribosomal DNA. J Dermatol. 26(5):276-81.

Moharram, A. M. and Abdel-Gawad, K. M. (1989) Keratinophilic fungi associated with rabbit claws
in Egypt. J Basic Microbiol. 29(7):437-40.

Moubahser, A. H., el-Naghy, M. A., Abdel-Fattah, H. M. and Maghazy, S. M. (1992) Keratinolytic


fungi in Egyptian soils. I. Baited with hair and wool. Zentralbl Mikrobiol.147(8):529-35.

Moubasher, A. A., Mazen, M. A., Moharram, A. M. and El-Shanawany, A. (1992) Clinical and
mycological studies of dermatophytic diseases in Egypt: I-Assiut Governorate. Proc.10th Ann. Sci.
Conf. April, 1992.

Omieczynski, D. T., Swatek, F. E. and Beaker, S. W. (1965) A rapid method for the isolation of
Coceidioides immitis from sputum. Health Lab, Sci. 2:35-39.

Orr, G.F. (1977) A new species of Gymnoascus. Mycotaxon 5:470474

Orr, G.F., Roy, K. and Ghosh, G.R. (1977) Gymnoascoideus, a new genus of the Gymnoascaceae.
Mycotaxon 5:459469.

Pappagianis, D. (1988) Epidemiology of coccidioidomycosis. Curr Top Med Mycol 2: 199-238.

20
Peterson, S. W. and Sigler, L. (1998) Molecular genetic variation in Emmonsia crescens and Emmonsia
parva, etiologic agents of adiaspiromycosis, and their phylogenetic relationship to Blastomyces
dermatitidis (Ajellomyces dermatitidis) and other systemic fungal pathogens. J Clin Micro
36:29182925.

Pugh, G. J. F. and Mathison, G. E. (1962) Studies on coastal fungi.III. An ecological survey of


keratinophilic fungi. Trans Brit Mycol Soc 45:567-572.

Quinet-Leimann, B.C., Vera-Pizzini, C., Medeiros-Muniz, M., Carvalho-Albuquerque, P., Fialho-


Monteiro, P. C., Santos-Reis, R., Almeida-Paes ,R., Santos-Lazra, M., Wanke B., Andrade-
Prez ,M. and Zancop-Oliveira RM. (2005) Histoplasmosis in a Brazilian center: clinical
forms and laboratory tests. Rev Iberoam Micol 22: 141-146.

Rippon, J. W. (1982) Host specificity in dermatophytoses. In: Baxter M (Ed.) Proceedings of the Eight
Congress of the International Society for Human and Animal Mycology. Massey University,
Palmerston North 28-33.

Saito, N. and Nei, M. (1987) The neighbour-joining method for reconstructing phylogenetic tree. Mol.
Biol. Evol. 4: 406-425.

Sandhu, G.S., Kline, B.C., Stockman, L., and Roberts, G.D. (1995) Molecular probes for diagnosis of
fungal infections. J. Clin. Microbiol. 33: 2913-2919.

Schutze G. E., Figin, R. D. (Ed.) and Cherry, J. D. (Ed.) (1998) Textbook of Pediatric Infectious
Diseases, 4th Edition: 2297 - 2303.

Scott, J. A, Malloch, D.W. and Gloer, J. B. (1993). Polytolypa, an undescribed genus in the
Onygenales. Mycologia 85:503508.

Sharma, R. and Rajak, R. C. (2003) Keratinophilic Fungi: Natures Keratin Degrading Machines!
Their Isolation, Identification and Ecological Role. Resonance 8: 28-40.

Shigeru Uchiyama, Seigo Kamiya and Shun-ichi Udagawa (1995) Five onygenalean fungi from
Japan. Mycoscience 36:211-220

Shukia , P., Shukla, C. B., Kango, N. and Shukla, A. (2003) Isolation and characterization of
adermatophyte, Microsporum gypseum from Poultary Farm Soil of Rewa (Madhya Pradesh),
India. Pak. J Boil. Sci., 6 (6):622-625.

Sigler, L (2002) The Onygenaceae and other fungi from the Order Onygenales. In: Howard DH, ed.
Pathogenic fungi in humans and animals. New York: Marcel Dekker

Sigler, L., Abbott, S. P. and Woodgyer, A. J. (1994) New records of nail and skin infection due to
Onychocola canadensis and description of its teleomorph Arachnomyces nodosetosus sp. nov. J.
Med. Vet. Mycol. 32:275285.

Sigler, L., Lumley, T. C., and Currah R. S. (2000) New species and records of saprophytic
ascomycetes (Myxotrichaceae) from decaying logs in the boreal forest. Mycoscience 41: 495-502

Singh, C. J. (2002) optimization of an extracellular protease of Chrysosporium Keratinophilum and its


potential in bioremediation of keratinic wastes. Mycopathologia. 156:151-156.

21
Sugiyama, M. and Mikawa, T. (2001) Phylogenetic analysis of the non-athogenic genus Spiromastix
(Onygenaceae) and related onygenalean taxa based on large subunit ribosomal DNA sequences.
Mycoscience 42:413421.

Sugiyama, M., Ohara, A. and Mikawa, T. (1999) Molecular phylogeny of onygenalean fungi based on
small subunit ribosomal DNA (SSU rDNA) sequences. Mycoscience 40:251258.

Sundaram, B. M. (1987) Incidence of keratinophilic fungi in rice-field soils Mycopathologia 97:43-44.

Swatek, F. E. (1956) Ecological and pathological investigations of Haplosporangium parvum and


Coceidioides immitis in Southern California. Doctoral thesis, University of California, Los
Angeles.

Thompson, J.D., Higgins, D.G., and Gibbson, T. D., (1994) CLUSTAL W. Improving the sensitivity of
progressive multiple sequence alignments through sequence wieghting. Position specific gap
penalties, and wieght matric choice. Nucleic Acid Res 22:2673-2680.

Udagawa, S. and Uchiyama, S. (1999) Taxonomic studies on new or critical fungi of non-pathogenic
Onygenales 1. Mycoscience 40: 277-290.

Udagawa, S. and Uchiyama, S. (2000) Taxonomic studies on new or critical fungi of non-pathogenic
Onygenales 3. Mycoscience 41:303-311.

Udagawa, S.. and Uchiyama, S. (2000) Acanthogymnomyces, a new genus of gymnothecial


Ascomycetes with setae and sulcate ascospores. Mycotaxon 14:411418.

Ulfig, K. (2000) The occurrence of keratinolytic fungi in waste and waste-contaminated habitats. In:
Biology f dermatophytes and other keratinophilic fungi, Kushwaha R.S.K. and Guarro, J. Revista
Iberoamericana de Micologa, Bilbao. pp. 44-50.

Ulfig, K., Paza, G., Worsztynowicz, A., Mako T., Tien, A.J. and Brigmon R. L. (2003)
Keratinolytic Fungi as Indicators of Hydrocarbon Contamination and Bioremediation Progress in
a Petroleum Refinery. Polish Journal of Environmental Studies Vol. 12, No. 2, 245-250.

Ulfig, K., Przysta, W. Paza, G., and Miksch, K. (2006) BIiodegradation of petroleum hydrocarbons
by keratinophilic fungi. soil and Water Pollution Monitoring, Protection and Remediation, 323.

Ulfig, K., Terakowski M., Paza, G. and Kosarewicz. (1996) . Mycopathologia 136, 41.

Untereiner, W. A., Scott, J. A., Naveau, F. A., Currah, R. S. and Bachewich, J. (2002) Phylogeny of
Ajellomyces, Polytolypa and Spiromastix (Onygenaceae) inferred from rDNA sequence and non-
molecular data. Stud Mycol 47:2535.

Untereiner, W. A., Scott, J. A., Naveau, F. A., Sigler, L., Bachewich, J. and Angus, A. (2004) The
Ajellomycetaceae, a new family of vertebrate-associated Onygenales. Mycologia, 96(4), pp. 812
821.

Van Oorschot, C. A. N. (1980) A revision of Chrysosporium and allied genera. Stud Mycol 20:189.

Vanbreuseghem, (1952) Technique biologique pour lisolement des dermatophytes du sol. Ann. Soc.
Belg. Trop. 32:173178.

22
Vanbreuseghem, (1952) Technique biologique pour lisolement des dermatophytes du sol. Ann. Soc.
Belg. Trop. 32:173178.

Vidal, P., Ulfig, K., Valmaseda, M. and Guarro, J. (1999) Studies on keratinophilic fungi. XI.
Chrysosporium undulatum sp. nov. Antonie Van Leeuwenhoek. 75(3):171-82.

Vidal, P., Vinuesa, M. A. , Sanchez-Puelles, J. M. and Guarro, J. (2000) Phylogeny of the


anamorphic genus Chrysosporium and related taxa based on rDNA internal transcribed spacer
sequences. Rev Iberoamericana de Micologa 17:2229.

Vidyasagar, G.M., Hosmani, N and Shivkumar, D. (2005) Keratinophilic fungi isolated from hospital
dust and soils of public places at Gulbarga, India. Mycopathologia 159: 1321.

Walch, H. A., Prbnow, J. F., Wyborney, V. J. and Walch, R. K. (1961) Coccidioidomycosis in San
Diego County and the involvement of transported topsoil in certain cases. Amer. Rev. Resp. Dis.
84: 359-363.

Warnock, D. W. (2000) Mycotic agents of human disease. In: Fleming DO, Hunt DL, eds. Biological
Safety. Principles and Practices. Washington, DC: ASM Press, 111120.

Weitzman, I., Chin, N. X., Kunjukunju, N. and Della-Latta, P. (1998) A survey of dermatophytes
isolated from human patients in the United States from 1993 to 1995. J Am Acad Dermatol. 39(2
Pt 1): 255-61.

White, T., Bruns, T., Lee, S. and Tayl J. (1990) Amplification and direct sequencing of
ribosomal RNA genes for phylogenetics, p. 315-322. In M. Innis, D. Gelfad, J.
Inc., New York, N. Y.

Woldeamanuel, Y., Mengistu, Y., Chryssanthou, E. and Petrini B. (2005) Dermatophytosis in


Tulugudu Island, Ethiopia. Med Mycol. 43(1):79-82.

Youssef, Y. A., el-Din, A. A., Hassanein, S. M. (1992) ccurrence of keratinolytic fungi and related
dermatophytes in soils in Cairo, Egypt. Zentralbl Mikrobiol. 147(1-2):80-5.

Zaki, S. M., Mikami, Y., El-Din, A. A., Youssef, Y. A. (2005) Keratinophilic fungi recovered from
muddy soil in Cairo vicinities, Egypt. Mycopathologia. 160(3):245-51.

23