Vol.2, No.11, 1247-1252 (2010) Natural Science
http://dx.doi.org/10.4236/ns.2010.211151
The occurrence of keratinophilic fungi in selected soils
of Ladakh (India)
Sunil K. Deshmukh*1, Shilpa A. Verekar1, Archana Shrivastav2
1Department of Natural Products, Piramal Life Sciences Limited, 1, Nirlon Complex, Off Western Express Highway, Near NSE
Complex, Goregaon (East), Mumbai India
2Department of Microbiology, College of Life Sciences, Cancer Hospital and Research Institute, Gwalior, India; *Corresponding
author:
Received 24 July 2010; revised 27 August 2010; accepted 30 August 2010.
ABSTRACT
138 soil samples were collected from various
loca-tions in Ladakh, a cold desert in the Hima-
layan region, India and the samples were
screened for the presence of keratinophilic fungi
using the hair baiting techniques. 58 isolates
were recovered and identified. The cultures were
identified based on their macro- and mi-
cro-morphological features. A total of six genera
and fourteen species were isolated namely
Amauroascus kuehnii (0.72 %), Aphanoascus
keratinophilus (4.34%), Aphanoascus terreus
(2.17%), Auxarthron alboluteum (0.72%), Aux-
arthron conjugatum (0.72%), Chrysosporium ar-
ticulatum (0.72%), Chrysosporium mephiticum
(0.72%), Chrysosporium minutisporosum (2.17%),
Chrysosporium siglerae (0.72%), Chrysosporium
sp. (1.44%), Chrysosporium tropicum (15.94 %),
Chrysosporium submersum (3.62%), Chryso-
sporium state of Ctenomyces serratus (6.52%)
and Geomyces pannorum (1.45%). The present
study shows that keratinophilic fungi exist in the
cold desert of Ladakh.
Keywords: Cold Desert; Ladakh; Soil Fungi;
Keratinophilic Fungi; India
1. INTRODUCTION
Ladakh, an interesting land deep within the folds of
the Karakoram mountain ranges, is also known as the
“land of passes” (La means pass and dakh means
land). Ladakh is bordered by the Karakoram chain of
mountains in the north and the Himalayas in the south.
In Ladakh, altitude ranges from about 2750m at Kar-
gil to 7672m at Saser Kangri in the Karakoram. The
temperature vacillates between 27C in summer to
–45C in winter. Ladakh is situated between latitudes
30°N-36°N and longitudes 76°E-79°E. The great Hi-
malayan range lying to the south forms a barrier to
monsoon in this area due to which Ladakh is an iso-
lated cold desert region. Due to longer winters, the
agriculture season is short with very little vegetation
providing wind breaks as cover and so, totally ex-
posed to the elements (e.g. wind, snow, rain, river,
valley etc.), it experiences high velocity dust storms
and snow blizzards. The region of Ladakh normally
remains land locked between October to June because
of snow and severe cold winter.
The climate and geographic diversity of Ladakh
make it a potential interesting area to study the distri-
bution of keratinophilic fungi. The objective of this
study was to report the occurrence of keratinophilic
fungi from selected soil habitats from various loca-
tions in Ladakh.
2. MATERIALS AND METHODS
One hundred and thirty eight soil samples were
collected during June-July 2004 and June-July 2005.
Sample collection was done only once in a year, as for
most of the year the land is covered with snow. The
samples were collected randomly from the superficial
layer (depth not exceeding 3-5 cm) with the help of
plastic spoon in sterilized polythene bags from vari-
ous areas of Ladakh in the state of Jammu and Kash-
mir, India (Table 1). During the collection, attention
was paid to the soils of uncultivated area, cultivated
area, Pasture, road side, glacier bank, river banks and
Pangong Tso. The soils are shallow and immature
containing large portion of mineral grains. The soils
are sandy with porous gravel and devoid of humus.
The pH of soil ranges from 6.5 to 8.5. The collected
samples were kept at 15C for maximum of two
weeks, if not processed immediately.
Keratinophilic fungi were isolated by the hair bait-
ing technique of Vanbreuseghem 1 using human hair

S. K. Deshmukh et al. / Natural Science 2 (2010) 1247-1252 1248
Table 1. Distribution of soil samples collected in Ladakh (India).
Sites Altitude (m)
No. of samples
Examined
No. of samples
Positive % Positive
Pangong Tso 4350 29 7 24.13
Chang La 5289 8 2 25.00
Durbuk 4100 7 2 28.57
Khardung La 5602 45 11 24.44
Tangste 4100 3 1 33.33
Lukung 4345 6 2 33.33
Nimmu 3154 18 14 77.77
Magnetic Hill 4267 5 3 60.00
Phey 3150 3 3 100.00
Leh 3505 9 8 88.88
Indus river 3505 5 5 100.00
Total 138 58 42.02
as keratin bait. For this, sterile Petri dishes half filled
with the soil samples and moistened with sterile tap
water were baited by burying sterile human hairs in
the soil. These dishes were incubated at room tem-
perature and examined daily from the fifth day for
fungal growth over a period of 4 weeks. After ob-
serving the growth under a stereoscopic binocular
microscope, isolates were cultured on Sabouraud’s
dextrose agar supplemented with chloramphenicol (50
mg/l) and cycloheximide (500 mg/l). These fungi
were identified based on the monographs of Sigler
and Carmichael 2, Oorchschot 3, Currah 4, von
Arx 5, Cano and Guarro 6 , Vidal et al. 7 , Sigler
et al. 8 , Sigler, Guarro and Punsola, 9 Cano and
Guarro, 10 using macro and micro-morphological
features of these cultures.
3. RESULTS AND DISCUSSIONS
Table1 indicates the occurrence of keratinophilic
fungi in the cold desert of Ladakh. The maximum
number of positive samples i.e. 42.02 % was recorded
from 138 soils. Out of soil samples collected 60.00 %,
77.77%, 29.62 %, 70.58%, 100%, 21.42% and 20.00%
samples are from uncultivated land, cultivated land,
pastures, road side, Indus river bank, Pangong Tso and
glacier banks respectively, were found positive for the
presence of keratinophilic fungi.
The results of the isolations are presented in Table
2. They reveal that out of 138 samples only 58 yielded
keratinophilic fungi. All 58 isolates were categorized
in 14 species of six genera namely Amauroascus
kuehnii (0.72%), Aphanoascus keratinophilus
(4.34%), Aphanoascus terreus (2.17%), Auxarthron
alboleuteum (0.72%), Auxarthron conjugatum
(0.72%), Chrysosporium articulatum (0.72%), Chry-
sosporium mephiticum (0.72%), Chrysosporium
minutisporosum (2.17%), Chrysosporium siglerae
(0.72%), Chrysosporium sp. (1.44 %), Chrysosporium
tropicum (15.94%), Chrysosporium submersum
(3.62%), Chrysosporium state of Ctenomyces serratus
(6.52%) and Geomyces pannorum (1.45%).
Chrysosporium tropicum was 15.94 % in distribu-
tion and was the most dominant species. It is a cos-
mopolitan species of Chrysosporium and has been
reported from various parts of India 11-13. Ch ry so-
sporium state of Ctenomyces serratus, 6.52 % was the
next most frequently isolated species. It was reported
from Indian soils by various workers 14,15. It is
interesting to note that Chrysosporium state of
Ctenomyces serratus was mostly isolated from culti-
vated soils. Aphanoascus keratinophilus, 4.34 %, was
the next most frequently isolated species followed by
Chrysosporium submersum, 3.62%. Aphanoascus
keratinophilus was reported from various parts of In-
dia 11,16, 17 . In this study, Chrysosporium submer-
sum is reported for the first time from Indian soils. It
was previously reported from river sediments in
Catalonia and from soil and dust samples collected
from Belgium [7]. In present study it is isolated from
the cultivated soils only.
The other fungi isolated were Chrysosporium ar-
ticulatum (0.72%), Chrysosporium siglerae (0.72%),
Chrysosporium mephiticum (0.72%), Chrysosporium
minutisporosum (2.17%) and Aphanoascus terreus
(2.17%). Aphanoascus terreus was reported from In-
dia’s plains by various workers and dominates the
mycobiota of Indian soils because it is adapted to
warmer conditions of India 11,1 3,18. But the per-
centage of occurrence of A. terreus is reported to be
much less in hilly areas of Jammu and Kashmir as
compared to Indian plain 11,19, which confirms our
findings. Various species of Chryso sp o rium have been
reported from Indian soils 11,18,20. Chrysosporium
articulatum is reported for the first time from India.
Chrysosporium mephiticum was reported from Indian

S. K. Deshmukh et al. / Natural Science 2 (2010) 1247-1252 1249
Table 2. Occurrence of keratinophilic fungi from cold desert of Ladakh ( India).
Source of soil samples Khardung La Chang
La Pangong TsoMagnetic hills Nimmu Indus river LehLukungPhey Tang- ste Durbuk Total % Distribution
Pastures
Gla-
cier
Road
side
Gla-
cier
Pas-
tures
Bank
of lake
Unculti-
vated soil
Cultivated
soil
River
bank
Road
side Pastures PasturesRoad Side Pastures
No.of samples
examined
23
17 5 8 15 14 5 18 5 9 6 3 3 7 138
No. of samples
positive 5 3 3 2 4 3 3 14 5 8 2 3 1 2 58
Distribution ( % ) 21.73 17.64 60.00 25.00 26.66 21.4260.0077.77 100.0088.8833.33100.0033.33 28.57 42.02
Fungi recorded
Amauroascus kuehnii - - - - - - - - - 1 - - - - 1 0.72
Aphanoascus kerati-
nophilus 3 - - - - 1 - 2 - - - - - - 6 4.34
Aphanoascus terreus - - 1 - - 1 - - - 1 - - - - 3 2.173
Auxarthron albolu-
teum - - - 1 - - - - - - - - - - 1 0.72
Auxarthron conjuga-
tum - - - - - 1 - - - - - - - - 1 0.72
Chrysosporium
articulatum - - - - - - - - - - - 1 - - 1 0.72
Chrysosporium
mephiticum - - - - - - - - - - 1 - - - 1 0.72
Chrysosporium
minutisporosum - - - - - - 1 1 - - 1 - - - 3 2.17
Chrysosporium
siglerae - - - - - - - - 1 - - - - - 1 0.72
Chrysosporium sp. - 1 1 - - - - - - - - - - - 2 1.44
Chrysosporium
tropicum 1 - 1 1 4 - 1 - 4 6 - 1 1 2 22 15.94
Chrysosporium
submersum - - - - - - - 5 - - - - - - 5 3.62
Chrysosporium state
of Ctenomyces
serratus
1 - - - - - 1 6 - - - 1 - - 9 6.52
Geomyces pannorum - 2 - - - - - - - - - - - - 2 1.45
Total 5 3 3 2 4 3 3 14 5 8 2 3 1 2 58
soils by Sigler et al. 9 and Deshmukh 21. In the
present study it is isolated from pasture soil of Lu-
kung. Chrysosporium minutisporosum is also reported
for the first time from Indian soils. Vidal et al. 7
recovered it for the first time from river sediments
(Fluvia, Muga, Ter, and Ebre rivers), Catalonia, Spain.
Labuda et al. 22 also recovered a single isolate from
the soil and children’s sandpit samples in city park of
Nitra and Nová Baňa (Slovakia). In present study this
fungi is isolated from soils of Lukung, Nimmu and
Magnetic hills.
Auxarthron alboluteum and Amauroascus kuehnii
were recovered from the soils of Chang La and Leh.
Auxarthron alboluteum has been reported as
Malbranchea albolutea from the soils of Utah 8 and
Amauroascus kuehnii was previously reported from
dung samples 4 and from soils of Lonar crater 23.
Auxarthron alboleuteum is reported for the first time
from India. The genus Malbranchea and its ana-
morphs is common soil inhabitant and includes
keratinophilic species which are potentially patho-
genic to man and animals. Keratinolytic Malbranchea
and its anamorphs have been reported from the other
parts of Indian by various workers 13,20,24,25.
Two isolates of unidentified species of Chrysospo-
rium were recorded from the glacier bank soils and
roadside soils of Khardung La and their systemic po-
sition is yet to be confirmed.
Geomyces pannorum was isolated from Khardung
La (1.44%). It is a psychrophilic fungus found ubiq-
uitously in temperate to Antarctic soils throughout the
world 26,27]. It is also reported from other parts of
India 28-30.
Auxarthron conjugatum was recorded from the
soils of Pangong Tso. It was also recorded from In-
dian soils 13,15,31 while surveying keratinophilic
fungi. The other species isolated from Pangong Tso
and its vicinity were Aphanoascus keratinophilus, A.
terreus, and Chrysosporium tropicum. The fungi iso-
lated from Pangong Tso shows their adaptation to
such conditions where there is no visible vegetation.
It also shows that these fungi are growing on the
keratinic material added to the lake by birds, grazing
and burrowing animals. In Ladakh, the higher altitude

S. K. Deshmukh et al. / Natural Science 2 (2010) 1247-1252 1250
pastures are actually more productive. Local people
appoint one or two members of the village to take all
the animals to high pastures. While there, the animals
graze the attendants spin wool, make butter and
cheese and collect dung for the winter.
Chrysosporium tropicum, Aphanoascus keratino-
philus, Geomyces pannorum and Chrysosporium state
of Ctenomyces serratus were isolated from Khardung
La and Auxarthron alboluteum, and C. tropicum from
Chang La. Isolation of these fungi at 5602 m to 5289
m shows that these fungi can survive at this height
and extremely low temperature (upto –40C). These
samples were collected from Military base at Chang
La and Khardung La, indicates that these fungi are
associated with human activity.
Ladakh, is a part of the state of Jammu and Kashmir.
Deshmukh 15 has reported Aphanoascus keratino-
philus, C. tropicum, Chrysosporium state of Ctenomy-
ces serratus, Geomyces pannorum, Malbranchea sp.,
Microsporum gypseum, M. nanum, M. van-
breuseghemii, Trichophyton ajelloi, T. terrestre and
Uncinocarpus reesii from glacier banks of Kashmir
(Srinagar, 1730 m; Gulmarg, 2650 m and Sonmarg,
2730 m altitude). In the present study the fungi viz. M.
gypseum, M. nanum, M. vanbreuseghemii, Trichophy-
ton ajelloi, T. terrestre and U. reesii were altogether
absent. Similarly while surveying the keratinophilic
fungi from Indian soils, Garg 11 had isolated Ar-
throderma quadrifidum, Aphanoascus keratinophilus,
A. terreus, Chrysosporium evolceanui, C. tropicum,
Chrysosporium state of Ctenomyces serratus, Tricho-
phyton ajelloi, T. mentagrophytes, Microsporum canis,
M cookie, and M. gypseum from soils of Srinagar,
Kashmir (1730 m altitude) of which A. quadrifidum, T.
ajelloi, T. mentagrophytes, M. canis, M cookie and M.
gypseum were altogether absent in the present survey.
Deshmukh and Verekar 32, had isolated Aphanoas-
cus keratinophilus, A. terreus, C. queenslandicum, C.
tropicum, Chrysosporium sp., C. xerophilum, Chryso-
sporium state of Ctenomyces serratus, Malbranchea
gypsea, Microsporum canis, Microsporum gypseum
and Trichophyton mentagrophytes from the western
Himalayan state of Himachal Pradesh (800 m to
3500m altitude) which is also adjacent to the state of
Jammu and Kashmir of which C. queenslandicum, C.
xerophilum, Mal. gypsea, M. canis, M. gypseum and T.
mentagrophytes were altogether absent in this study.
This may be due to harsh climate of cold desert or the
samples being collected from high altitude of 3100 to
5600 m and the complexity of the ecosystem.
Keratinophilic fungi play a vital role in nature in
the breaking down and mineralization of keratinous
substrate into simpler substances. Their ability to
grow on keratin regarded them as pathogens to human
and animals including livestock. For example, an in-
vasive infection was noted in an 18-year old woman
who was a bone marrow transplant recipient as Chry-
sosporium, where the infection began as a facial
swelling and extended into the central nervous system
33. Chrysosporium zonatum was reported causing
disseminated infection in a patient with chronic
granulomatous disease 34. In Japan, C. zonatum
strains were also isolated from bronchial lavage from
a female in Chiba and from a male in Kyushu. Both
patients were with pulmonary cavities 35. Gymnas-
cella hyalinospora was isolated from invasive pul-
monary infection in a patient with acute myceloge-
nous leukemia 36. Steininger et al 37 report the
case of brain abscesses by the Chrysosporium ana-
morph of Nannizziopsis vriesii in a 38-year-old,
HIV-seropositive Nigerian man. Geomyces pannorum
is also found as a causative agent of dermatomycosis
and onychomycosis in humans 38,39. Chrysospo-
rium ophiodiicola was isolated from a subcutaneous
granuloma of a snake (Elaphe obsolete obsoleta) 40.
The Chrysosporium anamorph of Nannizziopsis vrie-
sii has been isolated from cases of dermatitis in te-
nacled snakes 41, brown tree snakes 42, chame-
leons 43 , crocodiles 44, bearded dragons 45 and
from a nasal granuloma in an Ameiva lizard 46. In a
recent report, a Chrysosporium species related to
Nannizziopsis vriesii was isolated from a case of cu-
taneous hyalohyphomycosis from two green iguanas
47. Vissiennon et al 48 reported disseminated
mycotic alterations in skin, lungs and liver in male
garter snake (Thamnophis) infected with Chrysospo-
rium queenslandicum. Chrysosporium tropicum was
reported from comb lesion in two different breeds of
chicken in India 49. Thus, these fungi may be re-
garded as ‘‘opportunistic’’ pathogens.
4. CONCLUSIONS
In conclusion, our study indicates that keratino-
philic fungi exist in the cold desert of Ladakh. The
difference in the prevalence of keratinophilic fungi in
the soils of different parts of India may be attributed
to their tolerance and adaptation to various biotic and
abiotic factors such as ecological conditions, soil type,
and vegetation. Garg 11  also emphasized that the
climate and perhaps other environmental factors are
apparently important in determining the distribution
of keratinophilic fungi.
5. ACKNOWLEDGEMENTS
The authors are grateful to Dr. Somesh Sharma, MD, Piramal Life
Sciences, Mumbai and Dr. B. R. Shrivastav, Director, Cancer Hospital

S. K. Deshmukh et al. / Natural Science 2 (2010) 1247-1252 1251
and Research Institute, Gwalior for all kinds of support during the
work.
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