Fungi associated with the hairs of goat and sheep in libya.

The mycoflora on the hair in 25 samples of each of goats and sheep collected from Libya was analyzed using two isolation methods at 25℃. Seventy species and 3 varieties belonging to 31 genera were collected from the two substrates. The hairs of sheep were polluted with fungi than goat, contained high total counts and number of genera and species. Two species of true dermatophytes were isolated namely Trichophyton rubrum and T. terrestre. Several keratiophilic species were isolated of which Chrysosporium indicum, C. keratinophilum and C. tropicum were the most prevalent. The commonest saprophytes in order of frequency were members of the genera Aspergillus, Penicillium, Emericella, Alternaria and Cochliobolus.

Animals are known to carry dermatophytes and other keratinophilic fungi on their hairs. These animals may act as a source of human and animal infections by direct contact or by contaminating working areas and dwelling places (Ripon, 1982). Therefore, studies on dematophytes and keratinophilic fungi present on hair of domestic animals are of considerable significance.

The presence of keratinophilic fungi on hairs of various animals has been briefly reviewed by numerous researchers in many parts of the world (Rees, 1967; Gugnani et al., 1975; Aho, 1980, 1983; Lopez-Martinez et al., 1984; Bagy and Abdel-Hafez, 1985; Bagy, 1986; Ali-Shtayeh et al., 1988a, b, 2000; Kubo et al., 1990; El-Said and Abdel-Hafez, 1995; El-Said, 1996, 2002; Rashid et al., 1996; Camin et al., 1998; Gugnani, 2000; Hubalek, 2000; Alghalibi, 2001; Moses and Sunday, 2001; Shukia et al., 2003; Periasamy et al., 2004; Dobrowolska et al., 2006; Ulfig, 2007; Thanaa et al., 2008 and others).

This study aimed to isolate dermatophytes and keratinophilic fungi associated with the hair of sheep and goats in Libya.

Materials and Methods

A total of 50 hair samples of goats and sheep (25 each) were collected randomly from different localities in Jifarah region. The samples were placed in clean plastic bags and transferred to the laboratory. For isolation of fungi associated with the animals hair, hair-baiting technique and dilution-plate method were used.

Hair-baiting technique

For isolation of dermatophytes and other keratinophilic fungi, the hair-baiting technique (Vanbreuseghem, 1952) was employed. Five fragments from each sample were scattered on the surface of moistened sterile soil (20~25% moisture content) in sterile plates (2 plates for each sample). The plates were incubated at 25℃ for 10~12 weeks and the soil in plates was remoistened with sterile distilled water whenever necessary. The moulds which appear on the hair fragments were transferred to the surface of Sabouraud's dextrose agar medium (Moss and McQuown, 1969). The medium was supplemented with 0.5 g cycloheximide (actidione), 40 µg/ml streptomycin and 20 units/ml penicillin as bacteriostatic agents. The plates were incubated at 25℃ for 2~4 weeks and the developing fungal colonies were counted, identified and calculated per 10 hair fragments for each sample. The relative importance value (RIV) was calculated (Shearer and Webster, 1985; Ali-Shtayeh and Asad Al-Sheikh, 1988).

The dilution-plate method

For estimation of saprophytic fungi associated with the hair, the dilution-plate method as described by Johnson and Curl (1972) was used. Glucose-Czapek's agar medium was used in which rose-bengal (0.1 mg/ml) and chloramphenicol (500 µg/l) were added as bacteriostatic agents. Five plates were used for each sample and the plates were incubated at 25℃ for 2-3 weeks. The developing fungi were counted, identified and calculated per g hair.

Results and Discussion

Dermatophytic and keratinophilic fungi (using hair baiting technique at 25℃)

Eleven species belonging to 2 genera of dermatophytes and closely related fungi were isolated from goats (7 species and 2 genera) and sheep hairs (10 and 2). The most contaminated hairs were those of sheep with the highest total counts (288 isolates/250 fragments) and wide spectrum of species (10 species) than goat's hair (171 isolates and 7 species) as shown in Table 1.

Table 1

Total isolates (T I, calculated per 250 hair fragments), number of cases of isolation (NCI, out of 25 samples), occurrence remarks (OR) and relative importance values (RIV) of dermatophytic and keratinophilic fungi recovered from hairs of 25 animals of each goats and sheep at 25℃

Occurrence remarks (OR): H = high occurrence, between 13~25 (out of 25); M = moderate occurrence, from 7~12 cases; L = low occurrence, from 4~6 cases; R = rare occurrence, from 1~3 cases.

Chrysosporium was the most frequent genus and emerged in 92% and 96% of the samples comprising 91.2% and 87.8% of the total isolates and have RIV of 183.2 and 183.8 of goats and sheep, respectively. This genus was also, isolated from cloven-hooves and horns of goats and sheep, in Egypt, as reported by Abdel-Hafez et al. (1990). They observed that it was represented in 79% and 51% of the samples constituting 23.4% and 24.7% of the total fungi of goats and sheep, respectively. Ali-Shatayeh et al. (1988a) indicated that Chrysosporium was found in 9.3% of all Keratinophilic fungi on goat's hair from the West Bank of Jordan. Also, they isolated the above genus from different animals hair in Jordan and it was represented in 13.7% of cow, 14.6% of donkey, 15.8% of rabbit, 25.0% of cat and 50.1% of dog's hair. It was represented by 9 species of which C. indicum, C. keratinophilum and C. tropicum were the most prevalent species. They occurred in 28%, 36% and 48% of goat and 40%, 68% and 48% of sheep hairs comprising 6.4% and 15.9%, 21.6%, and 28.9% and 30.4% and 21.2% of the total isolates on the two substrates, respectively. These three species were also, predominant among fungi isolated from hairs of camel and goats (Bagy and Abdel-Hafez, 1985) and cloven-hooves and horns of goats and sheep (Abdel-Hafez et al., 1990). The above species were also, isolated from mammals in Venezuela by Moraes et al. (1967), Austrailia by Rees (1967), India by Gugnani et al.(1975), Jordan by Ali-Shtayeh et al. (1988b) and Egypt by Bagy (1986).

One species namely C. pannorum and an unidentified Chrysosporium species were of low frequency on goats hairs. They were encountered in 16% and 12% of the samples matching 10.5% and 5.2% and 14.6% and 7.3% of the total isolateds on goat, and sheep hairs, respectively. The previous identified species was found to be in 1.7% of goat hairs from Egypt (Bagy and Abdel-Hafez, 1985).

Some species were isolated only from one substrate and not from the other such as C. dermatitidis (8% of the samples and 4.7% of total fungi) from goat; C. asperatum (4% and 1.7%), C. carmichalii (8% and 2.4%) and C. queenslandicum (4% and 1.7%) from sheep hairs. Other less common fungi which were recovered from the two substrates included C. xerophilum (Table 1). Most of these fungi were recovered, with variable degrees and densities from animals hair or natural soil baited, with sterilized human or animals hair from Egypt (Bagy and Abdel-Hafez, 1985; Bagy, 1986; Abdel-Hafez, 1987; Moharram et al., 1988; Abdel-Gawad, 1989; Abdel-Hafez et al., 1989, 1990, 1991; Moharram and Abdel-Gawad, 1989; Abdel-Hafez and EL-Sharouny, 1990; El-Said, 1996, 2002; El-Said and Abdel-Hafez, 1995; Thanaa et al., 2008 and others) as well as from other parts of the world (Filipello Marchisio, 1986; Ali-Shtayeh, 1988; Ali-Shtayeh and Asa`d AL-Sheikh, 1988; Ali-Shtayeh et al., 1988a, b, 2000; Chabasse et al., 1989; Kubo et al., 1990; Rashid et al., 1996; Camin et al., 1998; Gugnani, 2000; Hubalek, 2000; Alghalibi, 2001; Moses and Sunday, 2001; Shukia et al., 2003; Periasamy et al., 2004; Dobrowolska et al., 2006; Ulfig, 2007 and others).

Dermatophytes on goat and sheep hairs were represented only by the genus Trichophyton. It occurred in 12% and 24% of the animal hair samples examined representing 8.8% and 12.2% of the total isolates and have RIV of 20, 7 and 36, 1 on goats and sheep, respectively (Table 1). This result is in agreement with the finding of Bagy and Abdel-Hafez (1985) who reported that this genus was in low occurrence on large mammals. Also, Abdel-Hafez et al. (1990) isolated this genus in rare frequency from cloven-hooves and horns of goats and sheep. It was represent by 2 species and 1 unidentified species of which T. terrestre was isolated from the two substrates in rare occurrence. It emerged in 8% of the samples constituting 2.9% and 3.5% of the total isolates recovered on goats and sheep, respectively. Otcenasek and Dvorak (1962) reported that T. terrestre was probably only a skin contaminant of small mammals from South Eastern Moravia. Aho (1983) isolated T. terrestre from different animals including goats. The other identified species namely T. rubrum (1.7% of total isolates) was isolated in rare frequency from sheep hairs and was not encountered from different substrates as reported by several researchers from all over the world (Abdel-Hafez et al., 1991; El-Said, 1996, 2002; El-Said and Abdel-Hafez, 1995; Ali-Shtayeh et al., 1988a, b, 2000; Alghalibi, 2001; Moses and Sunday, 2001; Shukia et al., 2003; Periasamy et al., 2004; Dobrowolska et al., 2006; Ulfig, 2006; Thanaa et al., 2008 and others).

Saprophytic fungi (on glucose-Czapek's agar at 25℃)

Fifty-nine species and 3 varieties appertaining to 29 genera were collected from 25 hair samples of each of goats (38 species + 3 varieties and 16 genera) and sheep (48 + 2 and 26) on glucose-Czapek's agar at 25℃ (Tables 2 and 3). Several saprobic and cycloheximide resistant fungi were encountered and the most prevalent species on the two substrates were members of Alternaria, Aspergillus, Emericella and Penicillium. These results are similar to those obtained by Aho (1980) who reported that members of Penicillium, Cladosporium, Asperillus, Alternaria, Scopulariopsis, Trichoderma and Trichothecium were the most common in order of frequency saprophytic fungi from hairs of domestic and laboratory animals in Finland.

Table 2

Number of species (NS), percentage count (%C, calculated per total fungi) and percentage frequency (%F, calculated per 25 samples) of various fungal genera recovered from hairs of 25 animals of each of goats and sheep on glucose-Czapek's agar at 25℃

Table 3

Average total count (ATC, calculated per g hair in all samples), number of cases of isolation (NCI, out of 25 samples) and occurrence remarks (OR) of fungal genera and species recovered from hairs of 25 animals of each of goats and sheep on glucose-Czapek's agar at 25℃

Occurrence remarks (OR): H = high occurrence, from 13~25 cases (out of 25); M= moderate occurrence, from 7~12 cases; L = low occurrence, from 4~6 cases; R = occurrence, from 1~3 cases.

Aspergillus was the most frequent saprophytic fungus emerging in all samples constituting 66.0% and 57.7% of total isolates on goats and sheep, respectively. From this genus 12 species and one variety were identified of which A. flavus, A. fumigatus, A. niger, A. ochraceus and A. terreus were the most predominant species. They occurred in 32~100% and 24~100% of the hairs examined comprising 2.8~16.8% and 1.2~27.8% of total fungi on the two types of substrates, respectively. A. candidus, A. carneus, A. terreus var. africanus and A. wentii were isolated only from goat hair but were not encountered on sheep hairs. The remaining Aspergillus species were less common (Table 3). Bagy and Abdel-Hafez (1985) reported that Aspergillus species was the second most frequent genus on the hairs of goat and camel from AL-Arish in Egypt. They noticed that A. niger, A. flavus, A. sydowii, A. fumigatus and A. nidulans were the most common species. Also, Moharram et al. (1988) they found noticed that Aspergillus (10 species +1 variety) was the first most dominant fungi on human hairs in Egypt.

Penicillium (9 species) was the second most predominant genus and occurred in 80% of the animal tested contributing 12.1% and 10.3% of the total moulds on the goats and sheep, respectively. P. chrysogenum, P. citrinum and P. puberulum were prevalent on the two substrates. They emerged in 20~40% and 20~56% of the samples having 1.8~3.5% and 1.3~3.1% of total fungi, respectively. P. duclauxii was isolated only from goat hairs while, P. aurantiogriseum and P. waksmanii from sheep hairs. The remaining 3 species of Penicillium were encountered on the two substrates in rare or low frequencies of occurrence (Table 3). Also, members of Penicillium were among the most common fungi on the hair of different animals from West Bank of Jordan (Ali-Shtayeh et al., 1988a, b), AL-Arish of Egypt (Bagy and Abdel-Hafez, 1985) and El-Bahrin (El-Said, 1996).

Alternaria (2 species), Cochliobolus (5) and Emericella (2 + 2 varieties) were also prevalent and isolated in high occurrence from one or two substrates. They were found in 20~64% and 20~72% of the samples tested comprising 1.8~6.8% and 1.2~9.1% of total isolates on goats and sheep, respectively. From the above genera, A. alternata, C. hawaiiensis, C. lunatus, E. nidulans, E. nidulans var. dentata and E. nidulans var. lata were common. Other species of the preceding genera were less frequent (Table 3). The three genera were isolated from the skin of dogs and cats (Bone and Hackson, 1971), cloven-hooves and horns of goats and sheep (Abdel-Hafez et al., 1990), hairs or goats, cows, donkeys and cats (Ali-Shtayeh et al., 1988a, b) and camel and goats hairs (Bagy and Abdel-Hafez, 1985).

The remaining genera were isolated in rare or low frequencies of occurrence and were encountered collectively in 7.6% and 18.7% of the total moulds on goat sheep hairs, respectively (Table 3).

In conclusion, analysis of the mycoflora of goat and sheep hairs indicated that there are several keratinophilic and saprophytic fungi on the animal hairs. The hairs of sheep were contaminated more than goat hairs. This may be due to the increasing of organic in sheep hair than goat hair. The amount of organic matter fluctuated between 0.340~0.899% in sheep and 0.191~0.694% in goat hairs. Some species were predominant on one substrate and less on the other.

Eleven species were encountered only from goat but not from sheep hairs. On the contrary, 23 species were isolated from sheep and not from goat hairs. Several investigators, in many parts of the world isolated the saprophytic fungi which were recovered in the present work from various substrates including foodstuffs; seeds; grains; root and leaf surface of numerous plants; hairs, claws, cloven-hooves and horns of domestic and wild animals; soils; muds and air.