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538 Journal of Applied Sciences Research, 8(1): 538-542, 2012 ISSN 1819-544X This is a refereed journal and all articles are professionally screened and reviewed ORIGINAL ARTICLES Prevalence of Trichophyton Violaceum Isolated From Clinical Samples In Ismailia Egypt 1 Marwa M. Azab, 1Nora F. Mahmoud, 1Salah Abd Allah, 2Alaa El Din. M.S. Hosny, 3Atef S. Shehata and 4Roshdy W. Mohamed 1 Department of Microbiology and Immunology, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt. Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt. Microbiology. 3 Department, Faculty of Medicine, Suez Canal University, Ismailia, Egypt. Dermatology. 4 Department, Faculty of Medicine, Suez Canal University, Ismailia, Egypt. 2 ABSTRACT The common cause of skin infections are dermatophytes and opportunistic fungi. Aim of this study was to isolate and identify Trichophyton Violaceum (T. violaceum) from clinical samples from patients with different mycoses. Clinical samples from 154 patients were subjected to potassium hydroxide (KOH) examination and culture isolation; causative agents were identified macroscopically and microscopically. One hundred and nineteen specimens of 154 samples were KOH positive (77.3%) and 93/154 (60.8%) samples were culture positive. Of these, highest isolation rate was obtained in dermatophytes were T. Violaceum which isolated in 33/154 (21.4%) specimens, among the patients suffering from dermatophytosis. The study signifies the importance of mycological examination in the diagnosis of various mycoses for their effective management and the prevalence of T. Violaceum among isolated dermatophytes. Key words: Clinical samples, dermatophytes, Trichophyton Violaceum, prevalence. Introduction Although fungi are world wide, only few of them are considered pathogenic. The pathogenic fungi may give rise to infections in animals and human beings. Most of the agents cause infection of the superficial layers of the integument and only very few give rise to systemic involvement. Recently there has been an increase in the incidence of fungal infections. This increase may be a result of frequent usage of antibiotics, immunosuppressive drugs and various conditions like organ transplantations, lymphomas, leukemia and human immunodeficiency virus (HIV) infections (Petmy et al., 2004). Dermatophytes are fungi capable of invading keratinized regions such as skin, hair, and nails of human beings and animals, causing diseases known as dermatophytoses (Jackson et al., 1999; Gra¨ser et al., 2000; Liu et al., 2000; Faggi et al., 2001; Frealle et al., 2007; Shehata et al., 2008). Skin infection due to dermatophytes has become a significant health problem affecting children, adolescents and adults. Several species of dermatophytes can cause dermatophytoses which need to be differentiated by culture studies. A correct diagnosis is important to initiate appropriate treatment and also essential for epidemiological purposes. In the background of immunosuppression, detection of these agents becomes mandatory for they effective management of mycoses to prevent further invasions. The present study was undertaken to isolate Trichophyton Violaceum causing mycoses among the patients attending El-Sheikh Zaid dermatology center, Ismailia governorate, which has got an average new outpatient turnover of 2000-3000 per year. Microscopic Examination: Direct microscopic examination was undertaken in 10% potassium hydroxide (KOH) wet mount for the specimens of skin scales, pus crust, while 20% KOH was employed for hair and nail specimens. Corresponding Author: Marwa M. Azab, Department of Microbiology and Immunology, Faculty of Pharmacy, Suez Canal University, Ismailia (41522), Egypt. Tel: (+2) 01224602602; E-mail: [email protected] 539 J. Appl. Sci. Res., 8(1): 538-542, 2012 Materials and Methods This study was undertaken for a period of one year from March 2010 to March 2011. All the clinically suspected 154 cases were subjected to mycological work up. The specimens included skin scales, hair, hair roots and pus in cases of superficial mycoses. Deep tissues were the specimens in deep mycoses. Culture Study: The KOH positive cases were subjected to culture study, scraping site was cleaned aseptically with 70% ethanol and the scales were collected in a sterile slide with the help of sterile scalpel. The culture was performed in two different sets of media; one sabouraud dextrose agar (SDA) with chloramphenicol 50 mg/L and cycloheximide 500 mg/L and the other was dermatophyte tested media (DTM). The culture tubes were incubated at 28°C and the culture growth was observed every two days and the tubes were discarded only after six weeks in the absence of growth. The mycological identification was based on macroscopic and microscopic examination of the culture isolates. The macroscopic examination of T. violaceum was characterized by duration of growth, surface morphology and pigment production on the reverse. Based on pigment production on the media, T. violaceum colonies are glabrous or waxy, wrinkled, heaped and deep violet or lavender in color. In addition, KOH preparation of hair specimen showing endothrix infection (Emmons et al., 1977). The microscopic examination of fungal growth was observed with lactophenol cotton blue stain. Nature of mycelium and conidia formation (macro and micro conidia) helped to differentiate this specie. Table 1: Distribution of different clinical forms among patients with dermatophyte infections. Clinical findings Clinical cases No. T. capitis 57 T. corporis 26 T. cruris 18 T. manuum 7 T. fasciei 13 T. pedis 16 T. unguium 10 T. barbae 4 T. imbricata 3 Total 154 T. = tinea % 37 16.9 11.7 4.5 8.4 10.4 6.5 2.6 2 100 Results and Discussion Among 93 positive culture dermatophytes (60.8%) out of 154 (21.4%) dermatophytoses cases, 33 isolates (35.5%) were T. violaceum. Specimens from all these cases were KOH positive, which isolated from different types of dermatophytoses lesions. Among T. violaceum 21(22.6%) isolates were obtained from scalp/scalp hair, 6(6.45%) isolates from body skin scales, 6(6.45%) isolates from face skin scales. Table 2: Frequency distribution of the isolated T. violaceum from patients according to clinical forms. Lesion Sites T. violaceum No. Percentage (%) T. capitis 21 22.6 T. Faciei 6 6.45 T. corporis 3 3.2 T. cruris 3 3.2 Total 33 100 T. = Tinea T. violaceum seemed to be the chief isolate from the scalp/ scalp hair (21/33). This agent is still the commonest isolate from cases of tinea capitis in Egypt. This agent was also isolated from 6 specimens of face skin scales (6/33). Six (6/33) isolates were isolated from both body and groin skin scales evenly. Of the total number of 154 collected specimens, only 119 isolates (77.3%) were KOH positive and 93/154 (60.8%) samples were culture positive. The higher load of the organism in the immunocompromized background could be the reason for such higher isolation rate. The isolation rate in this study seemed to be similar to various other studies where it has ranged from 60-88.4% (El-Garf, 1979; Abdel-Hafez et al., 1980; Abou-Eisha and El-attar1994; Hanan, 1998; Nermin, 2001; Nermeen, 2006; Shehata A.S. et al., 2008). More isolates could be obtained from scalp/ scalp hair compared to skin scales and the isolates were least from nail specimens. 540 J. Appl. Sci. Res., 8(1): 538-542, 2012 Table 3: Frequency distribution of T. violaceum culturally positive cases among the examined patients according to clinical forms, sex and age. sex Sex Male Female Age range (years) Total Clinical forms No. (%) No. (%) T. capitis 15 (45.5) 6 (18.2) 1-10 21 (63.6) T. fasciei 3 (9.1) 3 (9.1) 1-10 6 (18.2) T. cruris - (0) 3 (9.1) 50-60 3 (9.1) T. corporis - (0) 3(9.1) 1-10 3 (9.1) Grand Total 18 (54.5) 15(45.5) 1-60 33 (100) T. = Tinea, No. = Number Fig. 1: Morphological characters of isolated T. violaceum. Fig. 2: KOH preparation of skin scraping and hair specimen showing Endothrix type of hair invasion by T.violaceum. Fig. 3: Microscopy of T. violaceum showing broad, tortuous, much branched and distorted hyphae with irregular width and chains of asymmetrical chlamydospores. 541 J. Appl. Sci. Res., 8(1): 538-542, 2012 Discussion: T. violaceum seemed to be the chief isolate from the scalp/ scalp hair (21/33). This agent is still the commonest isolate from cases of T. capitis in Egypt, this similar to many other reports (Nasser, 1969; El-Garf, 1979; Abdel-Hafez et al., 1980; Fawzia, 1987; Ferial, 1987; Ibtisam, 1989; Nermin, 2001; Maysa, 2002; Mohammed, 2004). But dissimilar to that reported by (Hanaa, 1987; 1991, EI-Benhawi et al.; El-Attar, 1992; Fatti HI and Al-Samarai AM, 2000; Omar, 2004). T. violaceum also isolated from 12 specimens of skin scales (12/33). In the present study, males (54.5%) were more commonly affected than females (45.5%). This results were in agreement with that recorded by (El-Mazny, 1972; Mohammed, 2004; Ibtisam, 1989; Abdel-Hafez and El-Sharouny, 1990; Abou-Eisha and El-attar, 1994; Hanaa, 1987), in which the ratios were 73.7%:26.3%, 54% : 46%, 51% : 49%, 50% : 50%, 80% : 20% and 80% : 20% respectively. Male to female ratio was 1.78:1, which is disagree with other studies done by (Omar, 2000) reported that clinical lesions of T. capitis were found in 54.1% of children; most of them were girls. Male predominance may be due to increased outdoor physical activities and increased opportunity for exposure to infection than females. The present study shows that dermatophytosis was more common in the age group of 1-10 years (69 cases) followed by 31-40 years (26 cases), which is comparable with other studies done by (Hanaa, 1987; Hanan, 1998; Omar, 2000). All T. violaceum isolates were from patients aged 1-10, except three isolates were aged 5060 years. The highest incidence in youngest aged 1-10 years may be due to increased physical activity, poor hygiene and increased opportunity for exposure. The present study shows that mycological examination of causative agent is necessary to differentiate and treat dermatophytes cause dermatophytoses. 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