Diversity of keratinophilic fungi on human hairs and nails at four governorates in upper egypt.

The mycobiota of 160 hair and nail samples collected from 4 different governorates in upper Egypt were estimated using soil plate method for isolating keratinophilic and dermatophytic fungi. Twenty-three fungi were recorded on both hair and nail samples collected from the four governorates. Highest fungal diversity (20) was collected from Red Sea samples followed by Qena (18) and Aswan (17) while lowest fungal diversity was recorded from Sohage samples. The common genera were Aphanoascus, Aspergillus, Penicillium, Paecilomyces and Chrysosporium. The most prevalent species belonging to these genera were: A. fulvescens, Aphanoascus sp. A. flavus link, A. flavus var. columnaris, P. chrysogenium. P. lilacinus and C. sulfureum. True dermatophytes such as Nannizzia fulva appeared in 20~30% of the male samples.

The keratinophilic fungi are of prime importance in regard to various skin diseases prevalent in various areas. These fungi are able to utilize keratin, a fibrous protein, as sole carbon and nitrogen source and survive saprophytically in nature (English, 1963). Many keratinophilic fungi frequently parasitize keratinous tissue, viz. skin, nails and hair in man and animals. Some of them share certain morphological features, constituting aspecial group called dermatophytes (Gugnani, 2000). The impact of keratinophilic fungi on human health seems unexplored part of various studies (Shukia et al., 2003). Dermatophytes and other keratinophilic fungi have been isolated from various keratinized part of body in animals and human (Abdel-Hafez, 1987; Nicholis and Midgley, 1989; Ali-Shtayeh et al., 2000; Alghalibi, 2001; El-Said, 2002; Dobrowolska et al., 2006).

Various keratinophilic fungi along with some dermatophytes are responsible for various skin infections, little epidemiological data on the fungal disease of skin in human is available (Shukia et al., 2003). Knowledge of the frequency and extension of etiological agents of humans and animal mycosis and other potentially pathogenic fungi on the healthy hairs and nails of humans is of prime importance for understanding of epidemiological cycle of these fungi (Otcenasek, 1978; Lee et al., 1990).

The present work is aimed at evaluating the biodiversity of keratinophilic fungi in Upper Egypt among male and female students of South Valley University at Aswan, Qena, Sohage and Red Sea Governorates.

Materials and Methods

Sample collection

A total of 80 hair and 80 nail samples were collected from male and female students of South Valley University at Aswan, Qena, Sohage, and Red Sea Governorates. For the collection, 20 samples from each governorate and 10 samples sex were chosen at random. No samples taken from patients, all samples taken from students (aged 17~24 years). Hair and nail samples were placed, separately, in clean plastic bag and transferred to the laboratory and were stored at 4℃ until fungal analysis.

Sample analysis

The soil plate technique (Bagy et al., 1997; Efuntoye and Fashanu, 2001) was employed to estimate the diversity of keratinophilic fungi. Each sample (10 hair fragments or nails) scattered separately on surface of about 40 gm sterile Petri-dishes and the dust moistened with sterilized water (25~30 moisture content) and remoistened when ever necessary. Two plates were used for each sample and the plates were incubated at room temperature 4~5 weeks. Ten hair fragments (5 fragments from each plates) or ten nails (5 nails from each plates) were transferred to the surface of Sabouraud's dextrose agar medium (SDA) (Moss and McQuown, 1969) supplemented with antibiotics e.g. chloramphenicol (0.5mg/ml) and cycloheximide (0.5 mg/ml). Plates were incubated at 28 ± 2℃ for 10~21 days and the culture were examined periodically for fungal growth. After incubation isolates were cultured on SDA for identification on the basis of their colonial and morphological characters using monographic descriptions and other available literature (Stockdale, 1963; Van Oorschot, 1980; Spiewak, 1998; Kushwala, 2000).

Results

Keratinophilic fungi recovered from students' hairs

Thirteen population were recorded on 80 hair samples collected from four governorates. The highest fungal (11) was collected from Red Sea samples followed by Aswan (10) and Qena (9) while lowest, (8) was recorded from Sohage samples.

Dermatophytes and dematophyte-like keratinophilic fungi were represented by 3 genera of which Aphanoascus was the most frequent genus being recovered from 90~100% of both male and female students' hairs samples. It was represented by 2 species and 1 unidentified species of which A. fulvescences and Aphanoascus sp. were most common, emerging in 50~80%, 40~50%; 20~80%, 30~40%; 20~70, 50~80% and 40~50% and 30~50%, from male and female students' hair from Aswan, Qena, Sohag and Red Sea governorates, respectively. A. terreus was isolated only from female students' hair, occurring in 30~50% of the samples. Arthroderma curreyi also isolated only from female students' hair recovered from 10~30% of the samples from Qena and Red Sea Governorates. Nannizzia fulva recovered only from Qena male students' hair, emerging in 30% of the samples.

Other keratinophilic fungal species (4 genera) were also isolated, of which Aspergillus was isolated in high frequency of occurrence. It was isolated from 60~80% and 40~80% of the male and female samples, respectively. It was represented by 3 species and 1 variety, of which A. flavus and A. flavus var. columinaris were the most common. Paecilomyces was recovered in high frequency of occurrence from female samples, contributing in 20~80% of the total samples, but in low frequency (0~10%) from male students' hairs. Penicillium was isolated in low frequency of occurrence recovered from 10~30% and 0~20% of the samples, respectively. It was represented by 2 species, it were P. chrysogenium and P. funiculosum. Alternaria alternata was also isolated in low frequency of occurrence, occurring in 0~10% of both male and female samples.

Keratinophilic fungi recovered from students' nails

Twenty-one species and one variety belonging to 12 genera were recovered from male and female students' nails from Aswan, Qena, Sohage and Red Sea governorates.

Dermatophytes and closely related fungi were represented by 5 genera of which Aphanoascus was the most frequent genus and was recovered from 80~90% and 80~100% of both male and female students' nails from these governorates. It was represented by A. fulvescens and Aphanoascus sp. They were the most frequent species, they encountered in 50~60%, 50~40%; 40~70%, 30~40%, 40~50%, 80~90% and 70~80% and 30~0%, from male and female samples from the four Governorates. Chysosporium ranked second in frequency of occurrence which represented by 2 species of which C. sulfureum was isolated only from Sohage and Red Sea governorates. emerging in 20%, 0% and 40%, 20% of male and female nails samples. C. xerophilium was isolated only from Red sea male students' nails, contributing in 40% of the studied samples. Arthroderma curreyi, Myceliophthora vellerea and Nannizzia fulva were isolated only from Red sea samples of which Arthroderma, emerging 30% from female students' nails only. Myceliophthora and Nannizzia were recovered only from male samples encountered in 10% and 20%, respectively.

Other keratinophilic fungi were represented by 7 genera of which Aspergillus was isolated in high frequency of occurrence, emerging in 80~100% of both male and female students' nails from these governorates. It was represented by 6 species and 1 variety of which A. flavus and A. flavus var. columinaris were the most common. Penicillium ranked secondly among the most common from the other keratinophilic fungi genera. It was recovered from 10~80% and 20~30% of the samples. It was represented by 3 species of which P. chrysogenium was the most frequent. Gibberella represented by (G. pulcaris) ranked third after Penicillium. It was isolated from 10~30% and 10~40% of male and female samples, respectively. The remaining genera were recovered, but with low frequencies of occurrence from male and female students' nails and these were; Candida (C. albicans), Mucor (M. hiemalis) and Paecilomyces (P.lilacinus).

Discussion

Fungi associated with students' hairs

Thirteen fungi were collected from male and female students' hairs. Aphanoascus was the most common genus from male and female students' hairs. It was represented by A. fulvescens (anamorph: Chrysosporium keratinophilium), A. terreus (anamorph: C. indicum) and Aphanoascus sp. (anamorph: C. tropicum). Aphanoascus spp. were recorded from human hairs in Assiut, Egypt (Moharam et al., 1988) and human hairs at Qena and Red Sea governorates (Maghraby, 1994) and from hair and scalp of school children in Palestine (Ali-Shtayeh, 2000).

Arthroderma curreyi (anamorph: Chrysosporium anamorph of Artroderma curreyi) emerged in 10~30% only from female students' hairs at Qena and Red Sea. This species has of world-wide distribution (El-Said, 1995; Ulfig et al., 1996: Hubalek, 2000).

Nannizzia fulva (anamorph: Microsporium gypseum) ecountered in 30% only from Qena male students' hairs. Maghraby (1994) found M. gypseum (9.1%) from students' tinea capitis samples at Qena and Red Sea Governorates.

Member of Aspegillus (3 species + 1 variety), Paecilomyces (1), Penicillium (2) and Aternaria (1) were recovered from hairs of both sexs. These fungi were isolated previously from hairs of large mammals or feathers of poultry and human (Aho, 1983; Abdel-Hafez, 1987, 1990; Ali-Shtayeh, 2000; Alghalibi, 2001).

Fungi associated with students' nails

Twenty-one fungi belonging to 12 genera were recovered from male and female students' nails. Dermatophytes and related fungi were represented by 5 genera of which Aphanoascus was the most frequent genus. A. flvescens and Aphanoascus sp. were infrequently encountered from male and female nails. Previously, these species were isolated with different incidences, from students' nails (Abdel-Hafez and El-Sharouny, 1990; Abdel-Raouf, 2000), chicken and pigeon claws (Abdel-Hafez, 1989). Chrysosporium ranked second in frequency, it was represented C. sulfureum and C. xerophilium which was isolated previously from students' nails at Sohage Region (Abdel-Raouf, 2000).

Artroderma curreyi (Chrysosporium: anamorph Arthroderma curreyi), Myceliophthora verllera, and Nannizzia fulva (anamorph: Microsporium gypseum) were encountered through our study, those species were previously isolated by Katiyar and Kushwaha (2000) and Efuntoye and Fashanu (2001).

Several saprobic and cycloheximide resistant fungi such as Alternaria, Aspergillus, Candida, Gibberella, Mucor, Paecilomyces and Penicillium were recovered from students' nails. These species were isolated previously, but with different frequency, from students' nails (Abdel-Hafez and El-Sharouny 1990), cloven-hooves and horns of goats and sheep (Abdel-Hafez et al., 1990) and from domestic birds nails (Efountoye and Fashanu, 2001).

In conclusion, highest fungal diversity (20 species) was recorded from Red sea samples followed by Qena (18) and Aswan (17) while the lowest fungal diversity (16) was recorded from Sohage samples. Red sea governorates is coastal city with high humidity level while Qena and Aswan have higher temperature range than Sohage governorate. It is well known that high temperature and humidity are favored for fungal growth. High number of fungal species was recorded from nail samples compare to those recorded from hair samples at the four governorates and this is in agreement with the previous studies. (Moharam et al., 1988; Maghraby, 1994; El-Said et al., 1995; Abdel-Raouf, 2000; Ali-Shtayeh, 2000).

Table 1

Number of cases of isolation (NCI) of keratinophilic fungi isolated from nails and hairs of students of South Valley University at Aswan, Qena, Sohage and Red Sea Governorates

F = female, M = male, v = variety