DR Subclinical onychomycosis in patients with type II diabetes Amira Elbendary,1,2 Amira El Tawdy,1 Naglaa Zaki,1 Mostafa Alfishawy,3,4 Amr Rateb1 1Department of Dermatology, Kasr Al Ainy Faculty of Medicine, Cairo University, Egypt; 2Ackerman Academy of Dermatopathology, New York, NY, USA; 3Department of Internal Medicine, Kasr Al Ainy Faculty of Medicine, Cairo University, Egypt; 4Department of Medicine, Queens Hospital Center/Ichan school of Medicine at Mount Sinai, New York, NY, USA Abstract Fungal organisms could be present in the nail without any clinical manifestations. As onychomycosis in diabetics has more serious complications, early detection of such infec- tion could be helpful to prevent them. We aim in this study to assess the possibility of detect- ing subclinical onychomycosis in type II dia- betic patients and addressing possible associ- ated neuropathy. A cross sectional, observa- tional study included patients with type II dia- betes with normal big toe nail. All were sub- jected to nail clipping of the big toe nail, fol- lowed by staining with Hematoxylin and Eosin and Periodic-Acid-Schiff (PAS) stains and examined microscopically. A total of 106 patients were included, fungal infection was identified in eight specimens, all were uncon- trolled diabetes, and six had neuropathy. Using the nail clipping and microscopic examination with PAS stain to detect such subclinical infec- tion could be an applicable screening test for diabetic patients, for early detection and man- agement of onychomycosis. Introduction Reports about fungi that could be present in the nail without any clinical manifestations have been documented,1-4 and this was referred to as Subclinical onychomycosis.3 Evidence that these fungi could change from the passive form to induce superficial fungal infection in case there is defect in the immune system of the patient was reported.5 Onychomycosis is a well known complica- tion of diabetes mellitus. About one third of diabetic patients are affected.6 Although ony- chomycosis doesn’t represent a serious infec- tion in most people, its risk is increased in dia- betics, due to its limb threatening infection that could progress to ulcers and amputation as a result of the comorbidities present in dia- betics, namely peripheral neuropathy, macro and microvascular diseases and impaired immunity in addition to foot deformities,7 and thus it is critical to manage onychomycosis properly in such patients. The approach to diagnose onychomycosis could be painful, prolonged and complicated. Nail clipping is an easy doing procedure, pain- less, cheap and reasonable.8 Accordingly, the aim of our study was to investigate the presence of subclinical ony- chomycosis in diabetic patients using nail clip- ping as a diagnostic tool, and addressing pos- sible association of neuropathy with the occur- rence of subclinical infection. Materials and Methods A cross sectional, observational study included participants with type II diabetes mel- litus following up in endocrinology clinics pre- sented to the clinic by issues unrelated to ony- chomycosis. Patients with any clinical nail dys- trophy including discoloration, subungual debris, thickening, onycholysis, or patients with previous diagnosis of onychomycosis at least one year before the study were excluded. Age, sex and history of associated medical condition, diabetes related factors including type of diabetes, duration, associated peripher- al neuropathic symptoms (numbness, burning, tingling or loss of sensation) were recorded. Diabetic neuropathy was tested by testing the vibratory sensation by using a 128 Hz tuning fork on the interphalangeal joint of the right hallux comparing it to the dorsal wrist. Patients with lost vibration sense on the dorsal foot or feel stronger vibration on the wrist were considered having diabetic neuropathy. This method of detection of neuropathy was chosen due to its utility in clinical practice being simple and reliable.9 Heamoglobin A1c (HbA1c) was assessed for every patient; HbA1c level below 7.0% was con- sidered as controlled diabetic patients.10 Nail clipping was done from a normal big toenail for each patient. The clippings were subjected for Hematoxylin and Eosin stain (H&E) and Periodic-Acid-Schiff (PAS) stain- ing according to standard protocol. The histopathologic criteria to diagnose onychomycosis included the presence of parakeratosis, serous lakes, hyperkeratosis and inflammatory cells (neutrophils) that may suggest the possibility of onychomycosis in H&E stained specimens and the presence of hyphae invading the plate in PAS stained sec- tions. A total of 106 patients with type II diabetes were included in the current study, 20 males (18.9%), and 86 females (81.1%). The duration of diabetes ranged from 2 months to 20 years with mean duration (7.9) years. The patients’ age ranged from 25 to 75 years and a mean age Dermatology Reports 2015; volume 7:6099 Correspondence: Amira Elbendary, Ackerman Academy of Dermatopathology, 45E 32nd Street #9, New York, NY 10016, USA. Tel.: +1.212.889.6225. E-mail: aelbendary@residents.kasralainy.edu.eg Key words: Diabetes mellitus type II; onychomy- cosis: nail clipping: neuropathy: subclinical dis- ease. Contributions: the authors contributed equally. Conflict of interest: the authors declare no poten- tial conflict of interest. Received for publication: 5 July 2015. Accepted for publication: 7 July 2015. This work is licensed under a Creative Commons Attribution NonCommercial 3.0 License (CC BY- NC 3.0). ©Copyright A. Elbendary et al., 2015 Licensee PAGEPress, Italy Dermatology Reports 2015; 7:6099 doi:10.4081/dr.2015.6099 Table 1. Factors associated with patients found to be positive for subclinical onychomy- cosis. Factors, variables Total Patients with Percentage of these patients patients subclinical onychomycosis within same group Disease duration More than 10 years 42 5 7.8 Less than 10 years 64 3 7.1 Glycemic control Controlled 7 0 0 Uncontrolled 99 8 8.1 Neuropathy Present 48 6 12.5 Absent 58 2 3.4 [page 34] [Dermatology Reports 2015; 7:6099] No n c om me rci al us e o nly [Dermatology Reports 2015; 7:6099] [page 35] 51.2 (±9.2) years, 49 patients were less than 50 years. 99 patients were uncontrolled dia- betes (HbAlc�7) (93.4%), while the controlled were seven patients (6.6%). 43 patients were on oral hypoglycemic drugs (40.6%), and 63 were on insulin therapy (59.4%). Forty eight patients had neuropathy (45.30%) as revealed by absence or weak vibration sense using the tuning fork test. PAS stained specimens revealed eight cases positive for fungal infec- tion, in which uniform septate hyphae were found. This accounts 7.5% of the total patients. These PAS positive eight cases were five females and three males, with age ranged from 40 to 70 with mean age 51.6 (±8.7). Four patients were on oral hypoglycemic drugs (9.3% of patients on oral hypoglycemic drugs), and the other four patients were on Insulin (6.3% of patients taking Insulin). Factors that were found to be associated with patients with subclinical onychomycosis are demonstrated in Table 1. Although follow up was not intended in our study, we found that five out of the eight patients had associated tinea pedis and two patients developed clinical onychomycosis after two and three months respectively during their visit to dermatology clinic. Discussion and Conclusions The microscopic findings were not correlat- ed with culture, as the objective of this study was to search for subclinical infection in dia- betics, and addressing associated possible risk factors for it, regardless of the causative agent, and using the most accurate method reported in the literature (finding hyphae in PAS stained specimens obtained by nail clipping). Larger population of type II diabetic patients is needed to be investigated in further studies in order to answer the question: do we need to recommend screening diabetics for subclinical onychomycosis using nail clipping as a diagnos- tic tool in their routine checkup? Scant data about subclinical onychomycosis were reported (Table 2) and might be a finding in a percent of type II diabetic patients espe- cially the uncontrolled ones which can be asso- ciated with tinea pedis and neuropathy. Using the nail clipping and microscopic examination with PAS stain to detect such subclinical infec- tion could be an applicable screening test for diabetic patients, being non painful, reason- able, and simple test, aiming for early detec- tion and management of onychomycosis and hence decreasing the incidence of its possible serious complications in diabetics and perhaps the serious side effects of prolonged systemic antifungal treatment. References 1. Davies RR. Mycological tests and ony- chomycosis. J Clin Pathol 1968;21:729-30. 2. Baran R, Badillet G. Primary onycholysis of the big toenails: a review of 113 cases. Br J Dermatol 1982;106:529-34. 3. Walling HW. Subclinical onychomycosis is associated with tinea pedis. Br J Dermatol 2009;161:746-9. 4. Shemer A1, Gupta AK, Farhi R, et al. When is onychomycosis onychomycosis? A cross- sectional study of fungi in normal-appear- ing nails.Br J Dermatol 2015;172:380-3. 5. Baran R, Badillet G. Is an ungual dermato- phyte necessarily pathogenic?. Ann Dermatol Venereol 1983;110:629-31. 6. Gupta AK, Konnikov N, MacDonald P, et al. Prevalence and epidemiology of toenail onychomycosis in diabetic subjects: a mul- ticenter survey. Br J Dermatol 1998;139: 665-71. 7. Nather A, Bee CS, Huak CY, et al. Epidemiology of diabetic foot problems and predictive factors for limb loss. J Diabetes Complications 2008;22:77-82. 8. Fillus Neto J, Tchornobay AM. How the nail clipping helps the dermatologist. Ann Bras Dermatol 2009;84:173-6. 9. Meijer JW, Smit AJ, Lefrandt JD, et al. Back to basics in diagnosing diabetic polyneu- ropathy with the tuning fork! Diabetes Care 2005;28:2201. 10. American Diabetes Association. Standards of medical care in diabetes 2014. Diabetes Care 2014;37:S14. Brief Report Table 2. Previous studies reporting fungal infection in apparently normal nail. Authors Patients Patients with Method used to diagnose Associated included subclinical infection fungal infection findings Davis1 1954 170 (8.7%) Direct microscopy (KOH) or culture Active onychomycosis in other toenails Baran and Badillet2 46 with normal toenails 7 (15%) KOH, PAS staining where Established T. rubrum toenail from total of 113 with onycholysis appropriate; fungal culture infection Baran and Badillet2 52 2 (4%) KOH, PAS staining where appropriate; fungal culture None Walling3 101 7 (1.5%) out of 66 Nail clipping with PAS staining None 6 out of 35 (17%) Confirmed tinea pedis Shemer et al.4 585 54 (9.2%) KOH None 23 (3.9%) Culture None 18 (3.1%) KOH and culture None Our study 106 8 (7.5%) Nail clipping with PAS staining Diabetes mellitus type II No n c om me rci al us e o nly