Keywords: Onychomycosis, nail samples, Clinical signs, Fusarium species, Jammu.

Transcription

1 American International Journal of Research in Formal, Applied & Natural Sciences Available online at ISSN (Print): , ISSN (Online): , ISSN (CD-ROM): AIJRFANS is a refereed, indexed, peer-reviewed, multidisciplinary and open access journal published by International Association of Scientific Innovation and Research (IASIR), USA (An Association Unifying the Sciences, Engineering, and Applied Research) Fusarial an unrecorded report from Jammu district (India) Ruchi Bhou*, Geeta Sumbali University of Jammu, B.R. Ambedkar Road, Jammu (India). Abstract: Fusarium species which are well known as plant pathogens and soil saprophytes have a worldwide distribution now emerged as major opportunistic fungal agents. Several species of Fusarium have reported to cause superficial and subcutaneous infections, such as and keratomycosis, in humans. To stress the need to identify correctly and institute appropriate antifungal therapy in newly emerging human fungal infectious agents. Repeated mycological sampling of the nails of the suspected fungal infection were processed as per the standard format including direct microscopy and fungal culture on Sabouraud's dextrose agar. We report eight cases of caused by five different species of Fusarium i.e Fusarium solani (3), Fusarium verticilloides (1), Fusarium proliferatum (1), Fusarium pallidoroseum (2) and Fusarium chlamydosporum (1) from district Jammu, J&K. These species were isolated from both toenail and fingernail infection in all cases. The clinical appearance of the affected nails was total and distal subungual hyperkeratosis followed by onycholysis with white, yellow and brown coloration. The eight cases reported here suggest that Fusarium spp. should be taken into consideration in the differential diagnosis of fungal nail infection. Keywords: Onychomycosis, nail samples, Clinical signs, Fusarium species, Jammu. I. INTRODUCTION The present paper explores the association of different species of Fusarium as causal agents of in Jammu district (India). Onychomycosis is defined as a chronic fungal infection of toenails and fingernails leading to the gradual degradation of nail unit including the nail plate, nail matrix and nail bed. It represents 18 to 40% of all onychopathies and 39% of all superficial mycotic infections (1,2).Frequently it is caused by dermatophytes but now non- dermatophytic moulds are known to account for 2-12% of nail infections(3).the most important non- dermatophytic moulds causing include species of Alternaria, Aspergillus, Cephalosporium, Fusarium, Nattrassia and Scopulariopsis (3-5). Fusarium species are commonly soil saprophytes and plant pathogens but some of them are able to cause diseases in insects, reptiles, turtles and can also cause a variety of human infections including and foot infections(6,7).fusarial infections are characterized by the occurrence of white superficial lesions, but recently deep tissue infection and disseminated infections have greatly increased in patients with immunosuppressive conditions(8).in addition, close contact with soil, walking barefooted or with sandals, frequenting swimming pools and condition of trauma are regarded as predisposing factors for the development of and other infections due to Fusarium species(9). During exploration of in Jammu district, five species of Fusarium detected as causal agents. Here we report the eight cases of caused by F. solani, F. verticilloides,f. pallidoroseum, F. proliferatum and F. chlamydosporum. II. CASE REPORTS Nail clippings and scrapings from suspected individuals were collected and were processed by preparing a wet mount of nail sample in 20% potassium hydroxide (KOH) in dimethyl sulphoxide (DMSO) and then counterstained by chlorazol black E to enhance the visualization of the fungus. All the samples were cultured irrespective of the negative or the positive direct microscopic examination on Sabouraud dextrose agar (SDA) medium supplemented with chloramphenicol and incubated at o C for 21 days.repeated mycological sampling was done to identify the causal agent correctly. Detection of different Fusarium species was done by growing them on potato sucrose agar medium and by studying their cultural and morphological AIJRFANS ; 2015, AIJRFANS All Rights Reserved Page 21

2 characters.identification was done by following Booth (1971). Identity of some of the Fusarium species was also confirmed from Agharkar Research Institute,Pune (India). A. Case 1,2 and 3 Pathology of case 1,2 and 3: Case 1 of was detected in a 35 year old women who noticed symptoms first on the nail plate of left big toe nail. The nail was thickened, deformed, white to pale, showing nail bed hyperkeratosis. All these symptoms suggest a case of distal lateral subungual. Case 2 of was detected in a 60 year old women who noticed symptoms first on the nail plate of left big toe nail. The nail was thickened, deformed, white to pale, showing nail bed hyperkeratosis. All these symptoms suggest a case of total dystrophic. Case 3 of was detected in a 62 year old man who noticed symptoms first on the nail plate of left big toe nail. The patient is immunosuppressant suffering from cancer. The nail was thickened, deformed, pale to brown, showing nail bed hyperkeratosis and onycholysis. All these symptoms suggest a case of total dystrophic. Mycology of case 1, 2 and 3 Direct microscopic examination of the nail samples was detected to be negative. However, the nail samples cultured on SDA medium yielded a non dermatophytic fungus,whose colonies on PSA attained a diameter of 4-5cm in 4 days, dense floccose, white to cream; reverse brown in colour. Conidiophores branched, monophialidic; microconidia abundant,ovoid to oblong,0-1 septate, measuring x μm; macroconidia abundant, thick walled, borne on short multibranched conidiophores,3 septate, measuring x μm, with pedicellate foot cell. Chlamydospores were present in culture after days of incubation.based on the above features, the isolate was identified as Fusarium solani in all the three cases(fig.1). B. Case 4 Pathology: Case 4 of was detected in a 16 year old male student who noticed symptoms first on the nail plate of left finger thumb. The nail was thickened, deformed, white to pale, showing nail bed hyperkeratosis. All these symptoms suggest a case of leukonychia striata. Mycology: Direct microscopic examination of the nail sample was negative, but the nail sample cultured on SDA medium yielded a non dermatophytic fungus,whose colonies on PSA attained a diameter of 3 cm in 4 days, dense and delicately floccose, white to violet in colour. Conidiophores were simple, lateral, formed on short lateral branches. Microconidia abundant, formed in chains, fusiform to clavate, occasionally one septate, measuring x μm. Macroconidia were not observed in this isolate. Based on the above features, the isolate was identified as Fusarium verticelloides( Fig.2) AIJRFANS ; 2015, AIJRFANS All Rights Reserved Page 22

3 C. Case 5 Pathology: Case 5 of was detected in a 45 year old man who noticed symptoms first on the nail plate of all fingers of left hand. Infection began from the distal end and then spreads to the lateral sides of the nail infecting the whole finger nails. The nails were thickened, deformed, hard, pale to brown in colour, showing nail bed hyperkeratosis and onycholysis. All these symptoms suggest a case of distal subungual. Mycology: Direct microscopic examination of the nail sample was negative, but the nail sample cultured on SDA medium yielded a non dermatophytic fungus,whose colonies on PSA attained a diameter of cm in 4 days, dense and delicately floccose, creamish peach in colour. Conidiophores were branched and polyphialidic. Microconidia borne in short chains on polyphialides, 0- septate with flat base, measuring x μm. Macroconidia sparse on branched conidiophores, 1-septate, cylindrical and fusiform, measuring x μm. Based on the above features, the isolate was identified as Fusarium proliferatum ( Fig.3) AIJRFANS ; 2015, AIJRFANS All Rights Reserved Page 23

4 D. Case 6 and 7 Pathology of case 6 and 7: Case 6 of was detected in a 19 year old male student who noticed symptoms first on the nail plate of big toe nail of left foot.the nail was thickened, deformed, hard, pale to brown in colour, showing nail bed hyperkeratosis but no onycholysis. All these symptoms suggest a case of distal subungual. Case 7 of was detected in a 15 year old male student who noticed symptoms first on the nail plate of thumb nail of left hand. The nail was thickened, deformed, hard, pale to white, showing nail bed hyperkeratosis. All these symptoms suggest a case of total dystrophic. Mycology of case 6 and 7:-Direct microscopic examination of the nail sample was negative, but the nail samples cultured on SDA medium yielded a non dermatophytic fungus,whose colonies on PSA coverd whole Petri plate in 5-7 days, floccose, white with peach tinge and ultimately buff brown; reverse brown with peach tinge. Conidiophores were loosely branched and polyphialidic. Microconidia borne in short chains on polyphialides, 0-septate, measuring x μm. Macroconidia sparse, formed from aerial mycelium, 3-5 septate, curved,apex pointed, measuring x μm. Chlamydospores present. Based on the above features, the isolate was identified as Fusarium pallidoroseum in both the cases (as shown in Fig.4) E. Case 8 Pathology: Case 8 of was detected in a 54 year old man who noticed symptoms on the nail plate of thumb nail of right hand.the nail was thickened, deformed, hard, pale to white in colour, showing superficial white lesions, nail bed hyperkeratosis followed by onycholysis suggest a case of distal subungual. Mycology: Direct microscopic examination of the nail sample was negative but the nail sample cultured on SDA medium yielded a non dermatophytic fungus,whose colonies on PSA covered whole Petri plate in 5-7 days, produced pinkish, floccose colonies. Microconidia one celled, clavate, oblong to fusiform, borne directly on short and narrow phialides on sympodially proliferating polyphialidic conidiophores measuring,7-11x μm,canoe-shaped macroconidia 2-5 septate, measuring x μm were present, Intercalary, smooth, brown-walled chlamydospores were abundantly produced which were mostly globose and formed in short chains, The colony reverse was faintly brown in the beginning but became dark brown as the culture aged due to the increasingly abundant, darkly pigmented chlamydospores.based on the above features, the isolate was identified as Fusarium chlamydosporum (Fig.5). AIJRFANS ; 2015, AIJRFANS All Rights Reserved Page 24

5 V. DISCUSSION This report shows that the non-dermatophytic moulds are an increasing cause of. The incidence of these infections is probably influenced by environmental conditions as case numbers are increasing in tropical and humid climatic regions[10-14,15,16].over the last three decades, an increasing number of nondermatophytic filamentous fungi have also been recognized as agents of skin infections in humans and animals producing lesions clinically similar to those caused by dermatophytes [22-25].Probably just like dermatophytes, some non-dermatophytic filamentous fungi can also degrade keratin by producing proteolytic enzymes including keratinase[26,27]. Data obtained so far has been tabulated and analysed. As shown in table 1, fusarial nail infection was found to be associated maximum with the farmers of this region. This is probably due to the fact that they are usually working with the soil, which is a reservoir of fusarial species. The second group consisted of the businessmen and students who probably were careless with respect to hygiene. The third occupational group of people that were suffering from consisted of the housewives who did menial household work. Persual of data given in table 1 shows that among the various age groups, people between the age of years showed highest frequency of. This may be due to the less immunity with the increasing age. In addition, it was found that it is more prevalent in males than in the females. It was also observed that among the toe and finger nails that were screened, both were equally prone to fungal infection. Probably this may be due to the fact that both come in direct contact with the soil. During the present investigation, F. solani was isolated in all the cases from toenails. Earlier studies also reported F. solani from toenails(28, 29). In addition, two types of were diagnosed from the reported eight cases. These included the total and distal lateral subungual. These two types of have been reported earlier also as the most frequent types(11,19). The objective of the present study is to alert the medical community about the relevance of these opportunistic Fusarium species, which have emerged as human infectious agents, emphasizing the importance of correct etiological identification, allowing for appropriate treatment. Table 1. Clinical and epidemiological characteristics of the eight cases infected by various species of Fusarium Case Sex Agee Occupationn Infection sites Clinical Signs Case1 female 35 Farmer Toenail Distal lateral subungual Case2 female 60 Housewife Toenail Total dystrophic Case3 Male 62 Farmer Toenail Total dystrophic Evolution Organism Period Isolated 2 years F. solani 3 years F. solani 3 years F. solani AIJRFANS ; 2015, AIJRFANS All Rights Reserved Page 25

6 Case4 Male 16 Student Fingernail Leukonychia Striata(A type ofdistal lateral subungual ) Case5 male 45 Farmer Fingernail Distal subungual Case6 male 19 Student Toenail Distal subungual Case7 male 15 Student Fingernail Total dystrophic Case 8 male 55 Farmer Fingernail Distal subungual 1 year F.verticilloides 2 years F.proliferatum 6 months F. pallidoroseum 1 year F. pallidoroseum 2 years F.chlamydosporum ACKNOWLEDGEMENT The authors are grateful to University Grants Commission (UGC), New Delhi for funding this project. REFERENCES 1. Achten G and Wanet-Rouard J. Onychomycosis in the laboratory. Mykosen 1978; 1: André A and Achten G. Onychomycosis. Int J Dermatol 1987; 26(8): Summerbell RC, Kane J, Krajden S. Onychomycosis, tinea pedis and tinea manum caused by non-dermatophytic filamentous fungi. Mycoses 1989; 32: Plomer-Niezgoda E, Szepietowski J, Baran E, et al. Fungi from the genus Alternaria isolated from skin lesions: retrospective study of the last 20 years. Mikol Lek 1995; 3: Glowacka A, Wasowska-Krolikowska K, Skoron-Kobos J, Kurnatowski M, Janniger CK. Childhood : Alternariosis of all ten fingernails. Cutis 1998; 62: Oyeka CA, Gugnani HC. Skin infections due to Hendersonula toruloidea Scytalidium hyalinum, Fusarium solani and dermatophytes in cement factory workers. J Mycol Med 1992; 2: Guarro J, Gene J. Fusarium infections,criteria for the identification of the responsible species. Mycoses 1992;35: Guarro J, Gené J. Opportunistic fusarial infections in humans. Eur J Clin Microbiol Infect Dis 1995; 14: Comparot S, Reboux G, van Landuyt H, Guetarni L, Barale. Fusarium solani: Un cas rebelle d intertrigo. J Micol Méd 1995; 5: Bonifaz A, Cruz-Agular P, Ponce RM. Onychomycosis by moulds. Eur J Dermatol 2007; 17: Gianni C, Cerri A, Crosti C. Non-dermatophytic. An underestimated entity? A study of 51 cases. Mycoses 2000; 43: Gupta M, Sharma LN, Kanga AK, Mahajan VK, Tegta GR. Onychomycosis: clinico-mycologic study of 130 patients from Himachal Pradesh, India. Indian J Dermatol Venereol Leprol 2007; 73: Romano C, Gianni C, Difonzo EM. Retrospective study of in Italy: Mycoses 2005; 48: Boukachabine K, Agoumi A. Les au Maroc. Ann Biol Clin 2005; 63: Escobar ML, Carmona-Fonseca J. Onychomycosis by common non dermatophyte moulds. Rev Iberoam Micol 2003; 20: Hilmiog lu-polat S, Metin DY, Inci R, et al. Non-dermatophytic molds as agents of in Izmir, Turkey a prospective study. Mycopathologia 2005; 160: Mügge C, Haustein UF, Nenoff P. Causative agents of a retrospective study. J Dtsch Dermatol Ges 2006; 4: Gupta AK, Baran R, Summerbell RC. Fusarium infections of the skin. Curr Opin Infect Diseases2000;13: Jandial S, Sumbali G. Fusarial among gardeners: A report of two cases. Indian J Dermatol Venereol Leprol 2012;78: P Godoy, et al. Onychomycosis Caused by Fusarium solani and Fusarium oxysporum In São Paulo, Brazil. Mycopathologia April 2004 ; Volume 157, Issue 3, pp Kwon-Chung KJ, Bennet JE. Medical Mycology. Philadelphia, Lea & Febiger1992; Kane J, Summerbell R, Sigler L, Krajden S, Land G. Laboratory Handbook of Dermatophytes. Belmont, Ca (USA), Star Publishers, 1997; Punithalingam E, Western JM. CMI Descriptions of pathogenic fungi and bacteria No.274. Hendersonula toruloidea.kew, UK: Commonwealth Mycological Institute, Moore, MK. Skin and nail infection by non-dermatophyic filamentous fungi.mycoses 1978; 21 (Suppl. 1): English MP. Destruction of hair by two species of Chrysosporium. Trans Brit Mycol Soc 1976; 5: Nigam N, Kushwaha RKS. Biodegradation of wool by Chrysosporium keratinophilum acting singly or in combination with other fungi.trans Mycol Soc Japan 1992; 33: : Merz WG, Karp JE, Hoagland M, et al. Diagnosis and successful treatment of fusariosis in the compromised host. J Infect Dis 1988; 158: Robertson MJ, Socinski MA, Soiffer RJ, et al. Successful treatment of disseminated Fusarium infection after autologous bone marrow transplantation for acute myeloid leukemia. Bone Narrow Transplant 1991; 8: Girmenia C, Arcese W, Micozzi A, et al. Onychomycosis as a possible origin of disseminated Fusarium solani infection in a patient with severe aplastic anemia. Clin Infect Dis 1992; 14: AIJRFANS ; 2015, AIJRFANS All Rights Reserved Page 26