In vitro antifungal susceptibility of dermatophyte strains causing tinea pedis and onychomycosis in patients with non‐insulin‐dependent diabetes mellitus: a case‐control study

Background The efficacy of antifungal treatment may be reduced and/or delayed in diabetic patients. To date, no study has investigated the in vitro antifungal susceptibility of dermatophytes in this patient group. Objective We aimed to determine the dermatophyte species causing tinea pedis and onychomycosis, and in vitro susceptibility of these dermatophytes to terbinafine, itraconazole, and fluconazole in patients with non‐insulin‐dependent diabetes mellitus. We compared the findings in diabetic patients with those in non‐diabetic individuals. Materials and methods One hundred patients with non‐insulin‐dependent diabetes mellitus and 100 otherwise healthy controls clinically suspected with tinea pedis and/or onychomycosis were included. Skin scrapings and/or nail clippings were taken and cultured on Sabouraud dextrose agar, mycobiotic agar, and dermatophyte test medium. In vitro antifungal susceptibility tests were carried out according to the Clinical and Laboratory Standards Institute (CLSI) M‐38P protocol with some modifications. Results Fifty‐seven samples of 54 diabetics and 50 samples of 50 controls grew dermatophytes. In both groups, Trichophyton rubrum was the most common isolate. Mean MIC values of terbinafine, itraconazole, and fluconazole for all of the isolated dermatophyte strains were similar in two groups (P > 0.05). The difference in mean MIC values of three antifungals for T. rubrum and T. mentagrophytes between two groups was not statistically significant (P > 0.05). Conclusions Dermatophyte types causing tinea pedis and onychomycosis, their frequency patterns, and in vitro activity of three antifungals against dermatophytes in diabetics are similar to the non‐diabetics. Terbinafine is the most active agent in vitro in both groups.     

Treatment of tinea unguium

The modes of spread of fungal infection on the nails are discussed stressing the essential importance of the knowledge of this mode for finding of effective treatment of ungual trichophytosis. On the basis of own observations and reports of other authors combined operative and pharmacological treatment of trichophytosis is discussed and its usefulness is confirmed by the results of treatment and investigations. In 90 patients treated with griseofulvin 88.9% cures were obtained. In the light of these results the methods suggested by other authors for treatment of trichophytosis without surgical removal of nail plates are discussed.     

皮炎外瓶霉临床和环境分离株对6种常用抗真菌药物的敏感性测定

目的 探讨临床分离的皮炎外瓶霉对6种常用抗真菌药物的敏感性。方法 参考美国临床实验室标准化研究所M27-A2方案测定特比萘芬（TRB）、伊曲康唑（ITC）、两性霉素B（AMB）、氟康唑（FLC）、伏立康唑（VRC）及卡泊芬净（CAS） 对16株皮炎外瓶霉的最低抑菌浓度（MIC）,同时以E-test法测定VRC、ITC和AMB对16株皮炎外瓶霉的MIC,并进一步测定每种药物的最低杀菌浓度（MFC）。在此基础上,观察TRB与ITC及VRC联合应用时对皮炎外瓶霉的抗菌作用。结果 微量液基稀释法测得TRB、VRC、ITC、AMB、FLC和CAS 对皮炎外瓶霉MIC范围分别为：0.125 ~ 0.25、0.25 ~ 0.5、2.0、2.0、16 ~ 32和16 ~ 64mg/L;TRB、VRC、ITC、AMB和FLC的MFC范围分别为：0.125 ~ 0.5、0.25 ~ 0.5、2.0 ~ 4.0、2.0 ~ 4.0和16 ~ 64 mg/L。E-test法测定VRC、ITC和AMB的MIC范围为：0.032 ~ 0.094、0.047 ~ 0.5和0.125 ~ 3.0 mg/L。TRB与ITC及VRC在体外联合应用对皮炎外瓶霉无协同效应。结论 皮炎外瓶霉在体外对TRB、ITC、AMB、VRC敏感,对FLC和CAS不敏感。     

Identification of dermatophyte species causing onychomycosis and tinea pedis by MALDI-TOF mass spectrometry

Abstract:  Identification of dermatophytes is currently performed based on morphological criteria and is increasingly supported by genomic sequence comparison. The present study evaluates an alternative based on the analysis of clinical fungal isolates by mass spectrometry. Samples originating from skin and nail were characterized morphologically and by sequencing the internal transcribed spacer 1 (ITS1), ITS2 and the 5.8S rDNA regions of the rDNA clusters. In a blind comparative study, samples were analyzed by matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF MS). The mass spectra were compared to a database comprising of the spectral data of reference strains by applying the saramis software package. All fungal isolates belonging to the taxa Trichophyton rubrum , T. interdigitale , T. tonsurans , Arthroderma benhamiae and Microsporum canis were correctly identified, irrespective of host origin and pathology. To test the robustness of the approach, four isolates were grown on five different media and analyzed. Although the resulting mass spectra varied in detail, a sufficient number of signals were conserved resulting in data sets exploitable for unequivocal species identification. Taken together, the usually widespread dermatophytes can be identified rapidly and reliably by mass spectrometry. Starting from pure cultures, MALDI-TOF MS analysis uses very simple sample preparation procedures, and a single analysis is performed within minutes. Costs for consumables as well as preparation time are considerably lower than for PCR analysis.     

几种抗真菌药物对马尔尼菲青霉的体外药敏试验

目的 观察马尔尼菲青霉（PM）广西野生银星竹鼠寄生株与临床人分离株对伏立康唑和几种常用抗真菌药物的敏感性。方法 采用美国临床实验室标准委员会（CLSI）M27-A2和M38-A方案中的微量稀释法,测定伏立康唑、伊曲康唑、特比萘芬、两性霉素B 及氟康唑对14株广西野生银星竹鼠PM寄生株与25株临床人PM分离株25 ℃菌丝相及37 ℃酵母相的最小抑菌浓度（MIC）。用两样本均数比较t检验比较PM寄生株与临床人PM分离株MIC的差异性,用配对t检验比较同一株菌在两种不同温度相下的MIC差异性。结果 伏立康唑、伊曲康唑、特比萘芬、两性霉素B、氟康唑对PM寄生株菌丝相的MIC分别为：0.0313 ～ 0.1250、0.1250 ～ 1.0000、0.0313 ～ 0.5000、0.2500 ～ 4.0000、2.0000 ～ 8.0000 mg/L;对PM寄生株酵母相的MIC分别为：0.0078 ～ 0.2500、0.0313 ～ 0.5000、0.0313 ～ 1.0000、0.2500 ～ 2.0000、1.0000 ～ 8.0000 mg/L;对PM临床人分离株菌丝相的MIC分别为：0.0313 ～ 0.2500、0.0625 ～ 1.0000、0.0313 ～ 1.0000、0.2500 ～ 4.0000、2.0000 ～ 32.0000 mg/L;对PM临床人分离株酵母相的MIC分别为：0.0039 ～ 0.2500、0.0313 ～ 0.5000、0.0313 ～ 2.0000、0.1250 ～ 2.0000、2.0000 ～ 16.0000 mg/L。5种抗真菌药物对广西野生银星竹鼠PM寄生株与临床人PM分离株菌丝相和酵母相均敏感。同一温度下伏立康唑对两种不同来源PM的MIC最低,其他药物的MIC依次为伊曲康唑、特比萘芬 < 两性霉素B < 氟康唑。同一药物在同一温度下对广西野生银星竹鼠PM寄生株与PM临床分离株的MIC无明显差异;伊曲康唑、两性霉素B、特比萘芬对同一菌株在菌丝相和酵母相下的MIC存在差异。结论 PM对伏立康唑的敏感性最高;来自于广西野生银星竹鼠的PM寄生株与临床人PM分离株对伏立康唑、伊曲康唑、特比萘芬、两性霉素B及氟康唑的MIC类似;菌相的改变可影响PM对伊曲康唑、两性霉素B、特比萘芬的敏感性。     

In vitro antifungal susceptibility testing of fungi in patients with onychomycosis

Onychomycosis is the most common nail disorder. To examine in vitro antifungal susceptibility of fungi among onychomycosis patients. The study included 68 patients with onychomycosis. Nail specimens were cultured on Sabouraud dextrose agar and Dermasel agar base‐media. Isolated fungi were subjected to antifungal susceptibility tests against terbinafine, itraconazole, fluconazole, and griseofulvin. Candida species (Candida spp.) were detected in 32.4% of the cases of candidal onychomycosis (n = 37), 23.5% of the cases of distal and lateral subungual onychomycosis (n = 17), and 21.4% of the cases of total dystrophic onychomycosis (n = 14). Candida spp. were sensitive to fluconazole in 73.5%, itraconazole in 58.8%, and terbinafine in 5.9% of the cases. Aspergillus spp. were sensitive to itraconazole in all cases, and terbinafine in 87.5% of cases. Penicillium spp. were sensitive to itraconazole and terbinafine in 88.9% and 77.8% of cases, respectively. Trichophyton spp. were sensitive to terbinafine and resistant to itraconazole. Microsporum spp. were sensitive to itraconazole and resistance to terbinafine. All isolated fungi were resistant to griseofulvin. An increasing proportion of Candida spp. was observed among patients with different clinical varieties of onychomycosis. Candida spp. were highly sensitive to fluconazole and a lesser extent to itraconazole.     

兰州地区手、足、甲癣1,442例真菌学检查报告

手、足、甲癣临床虽为多见,但由于诊断条件不一,其发病数的统计就难以准确严格诊断标准,则应配合做真菌学检查.     

A review of the epidemiology of tinea unguium in the community

Tinea unguium is a common, chronic fungal infection of the nails. Many epidemiological studies have looked at the frequency with which this condition is seen in hospital outpatients clinics or mycological laboratories along with other dermatomycoses. Only recently have studies begun to emerge looking at the prevalence of this condition in populations. Hospital and mycological laboratory-based studies give valuable information about tinea unguium prevalence in a particular clinic, but cannot be compared with other studies due to confounding factors inherent in the different people attending individual clinics. From population-based studies the prevalence of tinea unguium lies between 2 and 8%. Tinea unguium increases steadily with age. It is infrequent but definitely found in children. With the increasing life expectancy in the Western world the prevalence of tinea unguium is likely to increase further without adequate prevention and treatment.     

In vitro susceptibility of 120 strains of Neisseria gonorrhoeae isolated in Kyrghyzstan

BACKGROUND: The World Health Organization has established a worldwide program for gonococcal antimicrobial surveillance, but so far no data on gonococcal susceptibility in Central Asia are available. GOAL: The need for biological data on the susceptibility of Neisseria gonorrhoeae in Kyrghyzstan, to enable adaptation of the national treatment protocol for gonococcal infections, led Médecins Sans Frontières and Epicentre to conduct a survey in collaboration with the Alfred Fournier Institute in Paris and the health authorities in Bishkek. STUDY DESIGN: In vitro susceptibility of N gonorrhoeae strains was determined with use of the reference agar-plate dilution technique. RESULTS: Results for 11 antibiotics tested on 120 strains of gonococci showed a low proportion (11.7%) of penicillinase-producing N gonorrhoeae and high proportions of intermediate or resistant strains to the majority of the antibiotics tested, including fluoroquinolones (>or=25% of strains resistant). All the strains were susceptible to spectinomycin, and only two strains had decreased susceptibility to cefixime. CONCLUSION: The therapeutic choices available in Kyrghyzstan appear to be limited to cephalosporins and spectinomycin.     

复发性外阴阴道假丝酵母菌病FUNGUS-7药敏研究

目的明确复发性外阴阴道假丝酵母菌病(Recurrent Vulvovaginal candidiasis,RVVC)致病白假丝酵母菌对7种常用抗真菌药物的敏感性差异。方法以FUNGUS-7真菌药敏试剂盒对已确定的213株白假丝酵母菌致病菌株进行2种多烯类药物、4种唑类药物和5-氟胞嘧啶共七种抗真菌药的体外药物敏感试验;结果 RVVC组致病白假丝酵母菌对7种抗真菌药物的敏感性依次为:制霉菌素两性霉素B5-氟胞嘧啶益康唑咪康唑氟康唑伊曲康唑;VVC组致病白假丝酵母菌对7种抗真菌药物的敏感性依次为:两性霉素B制霉菌素5-氟胞嘧啶咪康唑益康唑氟康唑伊曲康唑;RVVC组对制霉菌素和咪康唑的敏感性显著高于VVC组(P0.05);结论 RVVC和VVC致病白假丝酵母菌对抗真菌药的敏感性大体一致,RVVC治疗首选制霉菌素,VVC治疗首选制霉菌素或两性霉素B。     

Current topics in diagnosis and treatment of tinea unguium in Japan

The number of patients with tinea unguium accounts for 0.5% to 2% of the total number of outpatients in Japan. The diagnosis of tinea unguium can be relatively easily made on the basis of clinical symptoms and direct microscopic findings. However onychomycosis due to fungi other than dermatophytes should always be suspected if no dermatophytes are cultured despite positive findings in direct microscopy. In such cases, it is necessary to repeat cultures and conduct histopathological evaluation using PAS-staining technique or enzyme antibody technique. At present, oral administration of griseofulvin and terbinafine are the treatments covered by health insurance in Japan. In the future, however, other orally active antifungal drugs such as itraconazole and fluconazole will become available for use by in-patients who do not respond to griseofulvin. It is essential to patiently continue treatment even if oral therapy is impossible, because other therapies such as ODT therapy using topical antifugal agents and urea ointment or abrading affected nails as much as possible with a file or dental drill may be effective without oral therapy.     

In vitro susceptibility testing of Pityrosporum ovale against antifungal, antiseborrheic and antipsoriatic agents

Objective The in vitro antimicrobial susceptibility of Pityrosporum ovale strains isolated from patients with seborrheic dermatitis was determined. Method Minimum inhibitory concentrations of a total of 11 agents - including true antifungal, antiseborrheic, and antipsoriatic drugs - were measured. Results Ketoconazole was the most effective of the tested antifungal agents against Pityrosporum ovale in vitro (minimum inhibitory concentration 0.1 μg/ml). The antiseborrheic agents zinc pyrithione and selenium disulfide showed a good in vitro efficacy against Pityrosporum with similar minimum inhibitory concentrations. Liquor carbonis detergens and dithranol were also able to inhibit growth of Pityrosporum ovale in vitro, but much higher concentrations were necessary. Conclusion The tested agents commonly used against seborrheic dermatitis might exert their efficacy, at least in part, due to inhibition of Pityrosporum ovale.     

Epidemiology of tinea pedis and toenail tinea unguium in worshippers in the mosques in Adana, Turkey

The aim of this study was to determine the prevalence of dermatomycosis and the risk factors in those who perform their prayers in the mosques in the city center of Adana Province. The diagnosis of dermatomycosis was made on the basis of direct microscopy and/or culture in addition to clinical findings. Among 461 subjects, 136 (29.5%) had tinea pedis, 23 (5.0%) had tinea unguium, and 21 (4.5%) had both infections simultaneously, with a resulting total of 180 (39.0%) cases of dermatomycosis. The causative agents by frequency were: Trichophyton rubrum (67.0%), T. mentagrophytes var. interdigitale (31.1%), and Candida albicans (1.9%). The prevalence of the foot dermatomycosis was found to be high among those who practiced ablution 3-5 times a day and did not dry their feet immediately, who used rubber shoes, and/or who shared their shoes. Pedal dermatophyte infection seems to be a major problem among the adult Muslim male population regularly attending mosques especially in 5th and 6th decades of life.     

念珠地丝菌对五种抗真菌药的体外敏感性及电镜观察

目的 探讨5种临床常见抗真菌药对念珠地丝菌的体外抑制作用,并观察用药后念珠地丝菌的形态学变化。方法 5种临床常见抗真菌药为特比萘芬、两性霉素B、氟胞嘧啶、氟康唑和伊曲康唑。参照美国国家临床试验标准委员会修订的M27?A2方案及相关文献,测定各药物对念珠地丝菌的MIC值。应用扫描电镜观察特比萘芬作用后念珠地丝菌的形态学变化。结果 5种药物对念珠地丝菌的MIC值依次为：特比萘芬0.01 μg/ml、两性霉素B 0.4 μg/ml、氟胞嘧啶2 μg/ml、氟康唑2.69 μg/ml、伊曲康唑 0.25 μg/ml。念珠地丝菌对特比萘芬、两性霉素B、氟胞嘧啶、氟康唑均敏感,对伊曲康唑为剂量依赖性敏感。扫描电镜可见,经特比萘芬作用后念珠地丝菌菌体表面出现粗糙、皱缩、不规则缺损及孔洞,甚至呈碎片状。结论 特比萘芬对念珠地丝菌的抗菌活性最显著,在一定范围内,特比萘芬浓度越高,念珠地丝菌被破坏的程度越严重。     

Clinical study of Dermatophyte Test Strip,an immunochromatographic method,to detect tinea unguium dermatophytes

The Dermatophyte Test Strip visualizes mycotic antigens by immunochromatography. It allows easy and fast detection of dermatophytes. A multicenter, single‐arm, comparative clinical study was designed to evaluate the capacity of Dermatophyte Test Strip to detect dermatophytes in suspected tinea unguium specimens in comparison with direct microscopy and polymerase chain reaction (PCR). Signed consent was obtained from 222 subjects and all subjects completed the study. With the Dermatophyte Test Strip, dermatophytes were detected in 201 of 222 (90.5%) specimens but not in 21 of 222 (9.5%) specimens. With direct microscopy, dermatophytes were detected in 170 of 222 (76.6%) specimens but not in 52 of 222 (23.4%). Of the 45 specimens that showed inconsistent results between the two methods, PCR gave further results for 40 specimens, of which 37 (92.5%) specimens were positive and three (7.5%) were negative for dermatophytes. The positive concordance rate, negative concordance rate and overall concordance rate between the Dermatophyte Test Strip and direct microscopy were 81.1%, 66.7% and 79.7%, respectively. When inconsistent results were corrected using the results of PCR, these rates were 97.5%, 71.4% and 95.0%, respectively. When five specimens that could not be tested by PCR because no piece for the PCR test was left were excluded from analysis, these rates were 99.0%, 78.9% and 97.2%, respectively. The present results indicate good detection capacity of the Dermatophyte Test Strip. The Dermatophyte Test Strip provides a reliable, convenient and quick method to test for tinea unguium.