Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 18 September 2018 doi:10.20944/preprints201807.0236.v2 CHARACTERIZATION OF KERATINOPHILIC FUNGAL SPECIES AND OTHER NON-DERMATOPHYTES IN HAIR AND NAIL SAMPLES IN RIYADH, SAUDI ARABIA Suaad S. Alwakeel Department of Biology, College of Science, Princess Nourah bint Abdulrahman University, P.O. Box 285876 , Riyadh 11323, Saudi Arabia Telephone: +966505204715 Email: <[email protected]> < [email protected]> ABSTRACT The presence of fungal species on skin and hair is a known finding in many mammalian species and humans are no exception. Superficial fungal infections are sometimes a chronic and recurring condition that affects approximately 10-20% of the world‟s population. However, most species that are isolated from humans tend to occur as co-existing flora. This study was conducted to determine the diversity of fungal species from the hair and nails of 24 workers in the central region of Saudi Arabia. Male workers from Riyadh, Saudi Arabia were recruited for this study and samples were obtained from their nails and hair for mycological analysis using Sabouraud‟s agar and sterile wet soil. A total of 26 species belonging to 19 fungal genera were isolated from the 24 hair samples. Chaetomium globosum was the most commonly isolated fungal species followed by Emericella nidulans, Cochliobolus neergaardii and Penicillium oxalicum. Three fungal species were isolated only from nail samples, namely, Alternaria alternata, Aureobasidium pullulans, and Penicillium chrysogenum. This study demonstrates the presence of numerous fungal species that are not previously described from hair and nails in Saudi Arabia. The ability of these fungi to grow on and degrade keratinaceous materials often facilitates their role to cause skin, hair and nail infections in workers and other persons subjected to fungal spores and hyphae. Keywords: keratinophilic fungi, non-dermatophytes, mycobiota, hair, nails. INTRODUCTION and skin debris of dead animals (Bisen and Tewari, 2015). The Keratinophilic fungi are Fungi with affinities to attack basically saprophytes but occasionally becomes keratinized tissue are called "Keratinophilic potentially pathogenic to man and animals. fungi". These fungi are present in the The pathogenic forms of fungi are known as environment of all over the world, specifically “dermatophytes” and are known to cause in keratin containing habitats where humans superficial cutaneous infections and animals are living. The biological function (dermatophytoses) of keratinized tissues of of keratinophilic fungi in the soil is the humans and animals. Such fungi have better degradation of keratinized materials such as growth at temperatures of 25-28˚C with warm hides, furs, hair, feather, claws, nails, horns 3 © 2018 by the author(s). Distributed under a Creative Commons CC BY license. Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 18 September 2018 doi:10.20944/preprints201807.0236.v2 and humid conditions. Infections by fungi are Arabia, very few studies are available about relatively common in tropical countries due to the specific species that cause these infections environmental, social and economic conditions. and even fewer exist describing the species (Al- Superficial fungal infections are often chronic Sogair et al., 1991 and Venugopal, 1992). In a and recurring affecting approximately 10-20% study conducted among patients in Central of the world‟s population is affected during Saudi Arabia, T. mentagrophytes, Candida spp. their lifetime (Abanmi et al., 2008). Fungal and Aspergillus spp. were found to be the most infections of the skin, hair and nails are likely isolated species causing onychomycosis acquired via direct contact of other people, (Abanmi et al., 2008). Another study conducted infected animals or fomites (Alsheikh, 2009). in 2009 among patients clinically diagnosed Dermatophytosis can either be caused by true with dermatophytic infections in an Eastern dermatophytes (Microsporum, Trichophyton province of Saudi Arabia showed a variety of and Epidermophyton), yeasts (Candida), or species including Epidermophyton floccosum, moulds (e.g., Aspergillus, Alternaria, and Trichophyton rubrum, Trichophyton Fusarium) (Sahin et al., 2004). schoenlenii, and the non-dermatophytes In various geographical locations, Candida albicans and Fusarium (Alsheikh, several studies have been conducted on 2009) . Reports from Madina, Saudi Arabia characterization of fungi isolated from human (Hanafy, 2012) revealed that the most hair and nails. In Turkey, T. rubrum, T. frequently isolated causal agents of cutaneous mentagrophytes, T. verrucosum and T. mycoses were Microsporum canis (15.4%), violaceum. Microsporum canis, M. gypseum, Trichophyton metagrophytes (11.7%), and Epidermophyton floccosum, were commonly Trichophyton violaceum (11%). Screening for isolated from the hair and nails of students keratinolytic activity showed that M. canis and (Metintas and Kiraz, 2004). In northern Egypt, T. verrucosum recorded the highest value. In the most prevalent species included Riyadh City, Khaled et al. (2015) showed that Aphanoascus, Aspergillus, Penicillium, Tinea capitis infection had the highest Paecilomyces and Chrysosporium (Gherbawy prevalence among the patients (22.3%) while et al., 2006). In Northeast India, T. rubrum, T. Tinea barbae had the lowest. The identified mentagrophytes and M. gypseum were also the dermatophyte isolates were Trichophyton common isolates (Sarma, 2007). In Northern violaceum, Trichophyton verrucosum, Greece, dermatophytes including Trichophyton Trichophyton rubrum, Trichophyton rubrum (53.9%), Trichophyton mentagrophytes mentagrophytes, Trichophyton schoenleinii, (17.6%), and Microsporum canis (22.5%) were Trichophyton concentricum, Microsporum the most common isolates (Nasr et al., 2016). canis, Microsporum audouinii and These studies demonstrate the wide variety of Epidermophyton floccosum. Non dermatophyte fungal species that exist as normal flora or fungi included 5 isolates from Aspergillus, 4 possibly as colonizing non-pathological isolates from Acremonium and 15 isolates from organisms. Candida. M. canis was the most common Although, dermatophytic infections species (25% of isolated dermatophytes). In are a commonly encountered problem in Saudi Hail region of Saudi Arabia, Moursi (2016) 4 Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 18 September 2018 doi:10.20944/preprints201807.0236.v2 conducted an epidemiological study of MATERIALS AND METHODS dermatophytic diseases and found that 1) Collection of hair and nail samples: Trichophyton spp., are the predominant (82.11 Twenty-four workers in Riyadh, Saudi % of cases) followed by Epidermophyton spp. Arabia were recruited as participants between (16.55 %) but Microsporum spp., were the January and March 2016. Their jobs included lowest (1.34%). Among Tricophyton spp., T. construction, menial work and operation of gas mentegrophytes was more prevalent than T. and petrol stations. Most of the recruited rubrum (68.55% and 31.45%, respectively). workers had spent at least a year in Saudi More recently, Moursi et al. (2018) reported Arabia. We informed the participants of the that dermatophytes are the major pathogens aim and objectives of the study and obtained causing onychomycosis in Hail region. The written informed consent. The study protocol prevalent yeast was represented by Candida was reviewed and approved by the Princess albicans whereas the common non- Nourah bint Abdulrahman University dermatophytic mold was A. niger. Research Ethics Committee IRB No. H-01- R- The heterogeneity of the distribution 059. Hair and nail samples were obtained from pattern of dermatophytes in different parts of each worker using sterile instruments and the world has been attributed to various collection bags. factors, including climate, lifestyle, and the 2) Mycological analysis of human prevalence of immunodeficiency diseases in the hair samples: community, as well as the reluctance of patients to seek treatment because of i- Plating on Sabouraud`s embarrassment or the minor nature of disease Dextrose Agar (SDA) unless the condition became sufficiently serious Hair samples were individually placed on to affect the quality of life (Al-Sogair et al., the surface of Sabouraud`s Dextrose agar 1991 and Enugopal, 1992). Even fewer studies (SDA) which contained (g/l) glucose, 20, have attempted to understand the peptone, 10, agar, 20 and chloramphenicol, 0.5 heterogeneity of the commensal fungi of the according to the procedure described by Ellis region due to their relative non-pathogenicity et al. (2007). Chloramphenicol was compared to the bacterial flora of the human incorporated into the medium to suppress body (Efuntoye and Fashanu, 2002). Hence, bacterial growth. Cultures were incubated at this study was conducted to determine the 28°C for 1-3 weeks during which the growing diversity and distribution of the commensal fungi were examined and identified. Pure mycobiota from people living in Saudi Arabia cultures of fungi were kept on slants containing in an attempt to characterize, classify and the same medium for preservation and revision document these species and further understand (Efuntoye and Fashanu, 2002). their biology. The ability of some species to ii- Plating on sterile wet soil utilize keratin as an energy source i.e. The hair baiting technique originally keratinophilic fungi may aid in furthering our described by Vanbreuseghem (1952) was understanding of the interactions that