Kalyani Jatoth, T Mangilal and Sunil Junapudi, Prevalence of and other Keratinophilic Fungi from soil of playgrounds and public parks of Hyderabad, India. J.Curr.Biotechnol., 2016, 4(6):1-5.

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Prevalence of Dermatophytes and other Keratinophilic Fungi from soil of play- grounds and public parks of Hyderabad, India.

Kalyani Jatoth1*, T Mangilal2 and Sunil Junapudi3

1Department of Microbiology, Chandralabs, IDA prasanthi nagar,kukatpally, Hyderabad Telangana, India. 2Department of Pharmacy, UCT, Osmania University, Hyderabad, Telangana, India. 3Department of Pharmaceutical Chemistry, Geethanjali college of Pharmacy, cherryal, keesara, Ranga Reddy District, Telangan, India.

ARTICLE INF O ABSTRACT

Article History: Dermatophytes, related keratinophilic fungi were isolated from isolated from different Received 30 May 2016 locations in Hyderabad (Public parks, playgrounds, Zoological park- in and around Received in revised form 10 June animal and Bird enclosures). These samples were screened by using hair baiting 2016 techniques for isolation. Out of a total 60 samples, 52 (86%) were positive for growth Accepted 20 June 2016 of keratinophilic fungi. Eleven genera and 19 species were isolated and identified, of Available online 30 June 2016 which indicum (33.33 %) the most predominant species was isolated followed by C.tropicum (28.33%), flavus (25%), gypseum (16.6%) and terrestre (11.6%). Garden soils, followed by playground Key words: soils were found to be the most suitable for fungal growth. Some of the other fungi Keratinophilic fungi, Dermatophytes, isolated were C.zonatum , Alternaria alternata, Aspergillus niger, Aspergillus terreus, Hyderabad soils, Chrysosporium indicum, Fusarium moniliforme,F.solani , fulvescens etc. To our knowledge, this hair baiting technique. appears to be the first report concerning the isolation of keratinolytic fungi M. canis and Trichophyton terrestre from soils of Hyderabad by hair baiting technique. Introduction and veterinary epidemiologists (Marcella and Mercantini, Keratinophilic fungi are significant economically and play 1986). Keratinolytic fungi are a group of microorganisms important function in natural degradation of keratinous that are able to decompose keratin remains in environment substrates. Soils that are rich in keratinous materials are and are pathogenic to humans and animals. These fungi the majority conducive for the growth and occurrence of exist in communities together with keratinophilic fungi keratinophilic fungi. Their sharing is changeable with the that have weaker affinity to keratin and utilize chiefly the environment and depends on different factors, such as products of its decomposition (Dominik and human and or animal presence. Keratinous substances Majchrowicz, 1964). based on their occurrence in natural which are important natural material, happening in nature habitat, kertainophilic fungi are divided into three mainly in the form of hairs, wools, feathers, horns, categories: Anthropophilic, when human beings are the hooves, nails, skin and other cornified appendages natural hosts. Zoophilic, when a variety of animals act as constitutes natural baits for these fungi (Khanam and natural hosts. Geophilic, when the soil is the natural Jain , 2002). More than a few researchers have studied habitat (Ali-Shtayeh, 1989; Neetu, 2011; Piontelli, 1990; soil mycoflora with value to keratinophilic fungi in the Vollekova, 1992). past few decades (Al-Musallam 1988; Augt, 1995; Caglar, 2006; Deshmukh, 2008; Garg, 1985; Min, 2011; Ulfig, 1995). Hence it will be significant to analyze and identify the Keratinophilic fungi include a variety of filamentous fungi, mycoflora of school playgrounds, public parks and Zoo mainly comprising hyphomycetes and several other in order to evaluate the presence of keratinophilic fungi taxonomic groups. Hyphomycetes include dermatophytes and dermatophytes in these environments. This paper, and a great variety of non dermatophytic filamentous reports on the prevalence of keratinophilic fungi and its fungi (Mukesh and Sharma, 2010).The majority of related dermatophytes in the soils of Hyderabad, India. dermatophytes can live saprophytically and every This would help us know, the distribution and occurrence keratinophilic fungi can be considered as a potential of dermatophytes and other keratinophilic fungi; it will pathogen. Dermatophytes cause human and animal also through light on the risk of human mycoses and thus have drawn the attention of medical in these regions.

*Corresponding author. Email address: [email protected] Mobile: +918897724262 1 Int.J.Curr.Biotechnol. Volume 4; Issue 6; June, 2016 Materials and Methods isolated. C.tropicum (28.33%) was the second highest, Collection of soil samples: followed by Aspergillus flavus (25%). In the present A Sum of 60 soil samples was isolated from different study, M. gypseum (16.6%), was the most common locations in Hyderabad (Public parks, playgrounds, isolated followed by T.terrestre (11.6%). Zoological park- in and around animal and Bird However M.canis was isolated only from garden soils of enclosures). Soil samples were composed in sterile Zoo Park. C. zonatum, T. mentagrophytes, Alternaria polyethene bags by scooping up to a depth of 2- 5 cms alternata and Fusarium oxysporum were also isolated with the help of sterile disposable spoon. Each bag was besides other dermatophytes and keratinophilic fungi tightly packed and labeled indicating the site of collection listed in (Table 2). Both the playgrounds (43 isolates) and date. These samples were brought to the laboratory and garden soils (63isolates) were found to be rich and processed immediately and stored at room reservoirs of keratinophilic fungi and dermatophytes. temperature for further studies. Birds and animal enclosures (in and around of enclosures) also showed 19 and 7 isolates respectively. Isolation of keratinophilic fungi Sterile Petri dishes were half filled with methodically Discussion homogenized soil samples. Hair baiting technique was Previously, varied soil habitats have been screened from used for isolating the fungi. The half filled soil samples different countries e.g Iran, Brazil, Kuwait Pakistan, and were moistened with sterile distilled water; the amount India indicating that these groups of fungi are distributed of water added varied from sample to sample, depending worldwide (Al-Musallam, 1995; Da Silva, 2008; Itisha, on the moisture content of the sample. It was then baited 2010; Irshad, 2007; Fozia, 2007). The most frequently with short strands of sterilized human hair. Strands of isolated keratinophilic fungi in this study were human hair were spread uniformly as baits on the surface Chrysosporium indicum, C. tropicum, A flavus, M. of moistened soil samples. The plates were then incubated gypseum and T. terrestre. The high prevalence of these at 28±2o C for a period of 3-4 weeks. Five replicates were fungi from these soils explain that, hair of human and set for each sample (Keyvan Pakshir et al, 2013). animals and feather from birds which come to the soil either as dead or dropped off , serves as substrates and Isolation and Identification are subjected to microbial decomposition. Keratinophilic The hair baited plates were frequently examined for any fungi play a significant role in the natural degradation of fungal growth. Samples of hair showing fungal growth keratinized residues (Irshad, 2007; Sharma, 2003). were cautiously and aseptically chosen up and inoculate Deshmukh and Abdul was obtained 641 isolates from on Sabourauds dextrose agar (SDA) supplement with 125 soil samples of parks from Uttar Pradesh indicating chloramphenicol (0.05 mg/ml) and cycloheximide (0.5 mg/ soils of park to be a rich source of many keratinophilic ml) to make sure the bacterial and saprophytic fungal fungi and dermatophytes tropicum and C. indicum has growth, respectively. Prior to the inoculation on SDA, also been reported from soils (Abdel and Zaki, 2008), the baits were observed microscopically on a spotless isolated C. indicum from fields,animal and birds slide under a covered glass either containing a drop of enclosures, animal hairs and birds feathers also reported sterilized distilled water or any other staining solution large number of keratinophilic fungi from primary schools for any mixed growth. If it exhibited mixed growth, a dilute and public parks of Madras city indicating these soils to suspension of the material was transferred on SDA Petri be a reservoir of diverse fungi (Ramesh and Hilda, 1999). dishes in triplicates. After an incubation of 24 hours, the In our study Aspergillus flavus accounted for the third single germinating spores with initial hyphal growth were postion (25%).It was previously reported as second removed using a sterilized long needle and transfer to dominant species in soils of Gorgan areas in Iran fresh slants of SDA medium. In this way, mixed cultures (Moallaei et al, 2006). Among the dermatophytes, M. were made pure. These fungi were examined gypseum(16.6%) was the most predominant fungi isolated microscopically, identified according to their macro and from all the soils screened . M. gypseum was isolated micro morphological individuality, following the manuals from 13 hospital dust samples and it causes proposed by (Cano, 1990; Van Oorschot, 1980; Vishnu and tinea capitis in humans and is also reported from Sharma, 2015) and with the help of Osmania University, cats, dogs and rodents (Singh et al, 2009). The other Hyderabad, Telangana, India. dermatophytes isolated were Trichophyton terrestre(11.6%), T. mentagrophytes (8.33%) and Results T.rubrum (5%) followed in decreasing order. T. A Total of 60 soil samples were screened for the presence mentagrophytes is reported as the causal agent of tinea of dermatophytes and keratinophilic fungi of which, 52 pedis, tinea corporis, tinea cruris and . T. soil samples (86%) were positive for fungal growth. It mentagrophytes has been isolated from soils of was observed that all the soil samples collected from publicpark in Mumbai. While both T. terrestre and T. garden of Zoo Park were positive followed by public Parks mentagrophytes has been recorded from hospital dust and play grounds (Table 1). of Kanpur (U.P) (Singh et al, 2009). Chyrsoporium Where isolated 132 samples of isolates, belonging to 11 zonatum and Alternaria alternata accounted for the genera and 19 species, from various soils of hyderabad. same prevalence in the present study. Aphanoascus Some soils samples however yielded mixed growth, while fulvescens (10%) was isolated from garden soils. Two others represented single species. Chrysosporium Fusarium species were isolated from this study. Recorded indicum, C. tropicum, Aspergillus flavus, Microsporum Fusarium species to be the most frequently isolated gypseum and Trichophyton terrestre were isolated potential pathogens from soils collected in parks, cleared frequently. M. canis and T. terrestre were isolated for the areas, paddocks, river and roadsides of New Zealand first time from soils of Hyderabad by hair baiting (Simpanya and Baxter, 1996). Scopulariopsis brevicaulis technique. Garden soils, followed by playground soils was isolated from gardens, birds and animal enclosures. were found to be the most suitable for fungal growth. Scopulariopsis sp, Curvularia lunata and Fusarium C.indicum (33.33%) was the most predominant species solani has been reported from soils of gardens and school of Jhansi and Jaipur, (India) (Garg, 1985; Mukesh, 2010).

Volume 4; Issue 6; June, 2016 Int.J.Curr.Biotechnol. 2 Table - 1. Percent prevalence of keratinophilic fungi and dermatophytes isolated from soil samples of various habitats.

Habitat University campus Public Park Zoo Total

Play ground Garden Play ground Garden Garden Birds Animals

No of soil 10 8 9 8 10 6 9 60 samples studied No of samples 9 7 8 6 8 5 9 52 positive Percentage of 90% 87% 88% 75% 80% 53% 100% 86% positive samples

Table - 2: Frequency occurrence of keratinophilic fungi and dermatophytes from different soils of Hyderabad (India).

Fungi Isolated University Public Park Zoo Total % campus value Play Garden Play Garden Garden Birds Animals ground ground Aphanoascus - - - 2 3 1 - 6 10% fulvescens Aspergillus niger - - - 3 - 2 1 6 10% A.flavus 2 3 4 3 3 - - 15 25% A.terreus - 1 - - - 2 - 3 05% Alternaria 2 1 1 1 2 - - 7 11.6% alternata Chrysosporium 5 4 3 2 4 2 - 20 33.33% indicum C.tropicum 5 2 5 2 - 2 1 17 28.33% C.zonatum - - - 2 3 - 2 7 11.6% Curvularia lunata 1 - - 1 1 - - 3 05% Fusarium - - 3 1 2 - - 6 10% oxysporum F.solani - - - 1 2 - - 3 05% Microsporum 1 - 3 2 2 1 1 10 16.6% gypseum M.canis - - - - 2 - - 2 3.33% Penicillium - - - - 1 2 - 3 05% chrysogenum Rhizopus 1 1 1 2 - 1 - 6 10% stolonifer Scopulariopsis - - - - - 2 1 3 05% brevicaulis Trichophyton 1 - 1 - 1 - - 3 05% rubrum T.mentagrophytes - - 1 1 1 2 - 5 8.33% T.terrestre 1 1 1 - 1 2 1 7 11.6%

3 Int.J.Curr.Biotechnol. Volume 4; Issue 6; June, 2016 sites around Shivamogga City, Karnataka, India. The fungi isolated in this study are reported to be either Int.J.Curr.Microbiol.App.Sci. 5(2): 344-358. well known agents of mycosis or have been recovered from human and animal lesions such as Microsporum Augt M.M., Bayo J., Larrondo M.A, and Calvo M., 1995. gypseum, Trichophyton rubrum, Geotricum candidum, Keratinophilic fungi from soil of Brittany, France. Aspergillus flavus, Fusarium oxysporum, Mycopathol. 129: 81-82. Chrysosporium .sp and others. The dermatophytes are transmitted by contact with infected hair, fomites Avasn K., Lakshmi A., Rao S.R., Hossain K., Chaitanya (clippers, brushes) or from the environment (spores in D.A., and Karuna K., 1985. Dermatophytes and other soil). Dogs and cats harbor many saprophytic moulds fungi associated with hair-scalp of Primary school and yeasts on their hair coats and skin. children in Visakhapatnam, India: A Case Study And Literature Review. The Internet J. Microbiology. 5( 2): 1- The most common of these fungi isolated were species 4. of Alternaria, Aspergillus, Cladosporium, Mucor, Penicillium and Rhizopus. Most of these saprophytic Caglar S., and Akgun A., 2006. Effects of vesicular - isolates represent transient contamination by fungi in arbuscular mycorrhizal (VAM) fungi on the seedling soil and airborne fungi and are potential pathogens growth of three Pistacia species. J. Environ. Biol. 27: 485- causing mycosis (Avasn, 1985; Chmel , 1972; Rizwana, 489 2012). Cano J., Guarro J., 1990. The Aphanoascus. Mycol. It is clear from our results that soils of Gardens and Res. 94: 355–377. playgrounds are ideal environment for the growth of keratinophilic fungi and dermatophytes, and this could Chmel L., Hasilikova A., Hrasko J., and Vlacilikova A, be attributed to the high organic debris and keratinous 1972. The influence of some ecological factors on substrates like hair and feathers from birds and animals keratinophilic fungi in the soil. Sabouraudia. 10(1): 26- and plant litter present in these soils. However the 34 isolation of fungi was not uniform as it depends on organic matter. Organic matter content of soils is one of Da Silva Pontes Z.B.V., and Oliveira A.C., 2008. the major factors affecting the presence of keratinophilic Dermatophytes from urban soils in João Pessoa, Paraíba- fungi in soils (Ashwathanarayana, 2016; Katarzyna Brazil. Revista Argentina de Microbiología. 40: 161-163 Goralska, 2015). The study clearly indicates the varied distribution of keratinophilic fungi and dermatophytes Deshmukh S.K., Mandeel Q.A., and Verekar S.A., 2008. in soils of Hyderabad Keratinophilic fungi from selected soils of Bahrain .Mycopathologia. 165:143-147. Conclusion Based on the result of the present concentrate obviously Dominik T., and Majchrowicz I., 1964. A trial for isolating show that exhibit narrow diversity of keratinophilc fungi keratinolytic and keratinophilic fungi from the soils of and dermatophytes which is illustrative to its hot and the cemeteries and forests of Szczecin. Ekologia Polska arid environment Seria A. 12: 79- 105. Fozia I., Suhail M., and Abro., 2007. Keratinophilic fungi Acknowledgements from the soil of district,Jamshoro, Sindh( Pakistan). Authors are thankful to Osmania University, Hyderabad, Pakistan J. Bot. 39(4): 1377-1382. Telangana, India. For providing Technical support in the form of instruments, encouragement and guidance during Garg A.P., Gandotra S., Mukerji K.G., and Pugh G.J.E., 1985. research work. Ecology of keratinophilic fungi. Proc. Indian Acad. Sci. 94: 149-163. References Abdel R.M., and Zaki S.M., 2008. Experimental Irshad H.S., Kazi Y.F., Zardari M., and Shar A.H., 2007. Pathogenicity and Molecular Characterization of an Isolation of Keratinophilic Fungi from Soil in Khairpur Environmental Isolate of Chrysosporium zonatum Al City, Sindh, Pakistan.Bangladesh J Microbiol. 24( 1): 79- Musallam and Tan (Family: Onygonaceae, Order: 80. Onygonales).Int. J. Agri. Biol. 10 (3):273-277. Itisha S., and Kushwaha R.K.S., 2010. Dermatophytes and Ali-Shtayeh M.S., and Arda H.M., 1989. Isolation of related Keratinophilic fungi in soil of parks and keratinophilic fungi from floor dust in Arab elementary agricultural fields of Uttar Pradesh (India). Indian J and preparatory schools in the west bank of Jordan. Dermatol. 55(3): 306–308. Mycopathol. 106: 5-11 Katarzyna Goralska., Piotr Kurnatowski., Joanna Al-Musallam A.A., 1988. Distribution of keratinophilic B³aszkowska., and Anna Wojcik., 2015. Soil of Recreational fungi in desert soil of Kuwait.Mycos. 32: 296-302. Areas as a Reservoir of Keratinolytic Mould Fungi and Dermatophytes Potentially Pathogenic for Humans. Pol. Al-Musallam A.A., Al-Zarban S.H.S., Al-Sane N.A., and J. Environ. Stud. 24 (3): 993-1002. Ahmed T.M.M, 1995. A report on the predominant occurrence of dermatophytes species in cultivated soil Keyvan Pakshir., Moosa Rahimi Ghiasi., Kamiar from Kuwait. Mycopathol. 130: 159-161. Zomorodian., and Ali Reza Gharavi., 2013. Isolation and Molecular Identification of Keratinophilic Fungi from Ashwathanarayana R., and Raja naika., 2016. Prevalence Public Parks Soil in Shiraz, Iran. BioMed Research of Keratinolytic Fungi Isolated from the Poultry waste International. 2013:1-5.

Volume 4; Issue 6; June, 2016 Int.J.Curr.Biotechnol. 4 Khanam S.J.P., and Jain P.C., 2002. Isolation of keratin Van Oorschot C.A.N., 1980. A revision of Chrysosporium degrading fungi from soil of Damoh, India. Asian J. and allied genera. CBS,Baarn Netherlands. Stud. Mycol. Microbiol. Biotechnol. Environ. Sci. 4: 251-254. 20:1-89 Vishnu Sharma., Tarun Kumar Kumawat., Anima Sharma., Min J.L., Park J.S., Chung H., Jun J.B., and Bang Y.J., Ruchi Seth., and Subhash Chandra., 2015. Distribution 2011. Distribution of Soil Keratinophilic Fungi Isolated and Prevalence of Dermatophytes in Semi-Arid Region in Summer Beaches of the East Sea in Korea. Kor J Med of India. Advances in Microbiology. 5:93-106. Mycol . 16 (2):44-50. 7. Vollekova A., 1992. Keratinofilne huby v styroch lesnych podach (in Czech). Biol. (Bratislava). 47: 477-482. Marcella R., and Mercantini R., 1986. Keratinophilic fungi isolated from soils of the Abruzzo National Park Italy.Mycopathologia. 94:97-107. Mini K.D., Mini K., Paul., and Jyothis Mathew., 2012. Screening of fungi isolated from poultry farm soil for keratinolytic activity. Advances in Applied Science Research. 3 (4):2073-2077

Moallaei H., Zaini F., Pihet M., Mahmoudi M., Hashemi J., 2006. Isolation of Keratinophilic Fungi from Soil Samples of Forests and Farm Yards. Iranian J. Publ. Health. 35(4):62-69. Mukesh S., and Sharma M., 2010. Incidence of dermatophytes and other keratinophilic fungi in the schools and college playground soils of Jaipur, (India). Afr Journ of Microbiol Research. 4(24): 2647-2654. Neetu J., and Sharma M., 2011. Distribution of dermatophytes and other related fungi in Jaipur city, with particular reference to soil pH. Mycosis. 54(1):52-58. Piontelli E., Toro S.M., and Casanova D., 1990. Latitudinal distribution of and related Hyphomycetes in soils of northern Chile between 18-34 south latitude. Boletin Micol. 5: 79-106. Sharma R., and Rajak R.C., 2003. Keratinophilic fungi. Nature’s keratin degrading machines. Their isolation identification and ecological role. Resonance. 2003: 28- 40 Simpanya M.F., and Baxter M., 1996. Isolation of fungi from soil using keratin-baiting technique. Mycopathologia.136(2): 85-89. Singh I., Mishra M., and Kushwaha R., 2009. Dermatophytes related keratinophilic and opportunistic fungi in indoor dust of houses and hospitals. Indian J Med Microbiol. 27:242-246. RameshV.M., and Hilda A., 1999. Incidence of keratinophilic fungi in the soil of primary schools and public parks of Madras City (India). Mycopathologia. 143: 139–145.

Rizwana H., Amal Abdulaziz Al Hazzani., and Siddiqui I., 2012. Prevalence of dermatophytes and other keratinophilic fungi from Soils of public parks and playgrounds of riyadh, saudi arabia. The Journal of Animal & Plant Sciences. 22(4): 948-953

Ulfig K., and Korcz M., 1995. Isolation of keratinolylic fungi from a coal mine dump.Mycopathol. 129: 83-86

Vanbreuseghem R., 1952. Technique biologique pour I’ isolement des dermatophytes dusol. Ann Soc Belge Med Trop. 32: 173-78.

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