Isolates Of Trichophyton Rubrum Research Articles

Antifungal Activity of Efinaconazole Compared with Fluconazole, Itraconazole, and Terbinafine Against Terbinafine- and Itraconazole-Resistant/Susceptible Clinical Isolates of Dermatophytes, Candida, and Molds.

Recently, an increasing number of resistant-to-terbinafine dermatophytosis cases have been reported. Thus, identifying an alternative antifungal agent that possesses broad-spectrum activity, including against resistant strains, is needed. We compared the antifungal activity of efinaconazole with that of fluconazole, itraconazole, and terbinafine against clinical isolates of dermatophytes, Candida, and molds using in vitro assays. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of each antifungal agent were quantified and compared. Susceptible and resistant clinical isolates of Trichophyton mentagrophytes (n = 16), Trichophyton rubrum (n = 43), Trichophyton tonsurans (n = 18), Trichophyton violaceum (n = 4), Candida albicans (n = 55), Candida auris (n = 30), Fusarium spp, Scedosporium spp, and Scopulariopsis spp (n = 15 for each) were tested. Efinaconazole was the most active antifungal agent tested against dermatophytes, with MIC50 and MIC90 (concentrations that inhibited 50% and 90% of strains tested, respectively) values of 0.002 and 0.03 µg/mL, respectively. Fluconazole, itraconazole, and terbinafine showed MIC50 and MIC90 values of 1 and 8 µg/mL, 0.03 and 0.25 µg/mL, and 0.03 and 16 µg/mL, respectively. Against Candida isolates, efinaconazole MIC50 and MIC90 values were 0.016 and 0.25 µg/mL, respectively, whereas fluconazole, itraconazole, and terbinafine had MIC50 and MIC90 values of 1 and 16 µg/mL, 0.25 and 0.5 µg/mL, and 2 and 8 µg/mL, respectively. Against various mold species, efinaconazole MIC values ranged from 0.016 to 2 µg/mL versus 0.5 to greater than 64 µg/mL for the comparators. Efinaconazole showed superior potent activity against a broad panel of susceptible and resistant dermatophyte, Candida, and mold isolates.

COMPARISON OF THE ANTIFUNGAL ACTIVITY OF BLACK, WHITE, AND GREEN TEA KOMBUCHA EXTRACTS AGAINST DERMATOPHYTE FUNGI

Kombucha is obtained by fermentation of tea and sugar by bacteria and yeast. This beverage has several beneficial effects, including antimicrobial activity. The objective of this study was to evaluate the antifungal activity of Kombucha extracts against dermatomycosis-causing fungi. Different types of Kombucha (black, white, green tea) and times of fermentation (10, 20, 30 days) were evaluated. Kombuchas were filtered, and extracts obtained by liquid-liquid extraction with ethyl acetate. Minimum inhibitory concentrations (MIC) were determined against clinical isolates of dermatophytes Trichophyton rubrum and T. mentagrophytes by broth microdilution. Toxicity tests and high-performance liquid chromatography (HPLC) were also performed. MIC ranged from 62.5–2000 μg/mL. Green tea Kombucha and 10-days fermentation extracts showed the best results. Toxicity was observed only for white tea Kombucha. Despite the chromatographic profiles, further studies are needed to characterize the active molecules. However, Kombucha extracts may constitute a viable alternative for new treatment strategies against dermatomycosis.

Activity of amorolfine or ciclopirox in combination with terbinafine against pathogenic fungi in onychomycosis-Results of an invitro investigation.

Onychomycoses are difficult-to-treat fungal infections with high relapse rates. Combining oral and topical antifungal drugs is associated with higher success rates. Additive or synergistic modes of action are expected to enhance treatment success rates. Investigation of the combined effects of antifungal drugs invitro with different modes of action and application on clinical isolates from mycotic nails. Isolates of Trichophyton rubrum, Trichophyton interdigitale and Scopulariopsis brevicaulis were collected from infected toenail specimens of patients with onychomycosis. Susceptibility testing was performed in 96-well polystyrene plates using a standard stepwise microdilution protocol. Additive or synergistic activity at varying concentrations was investigated by the checkerboard method. Combining terbinafine with amorolfine tended to be more effective than terbinafine in conjunction with ciclopirox. In most combinations, additive effects were observed. Synergy was detected in combinations with involving amorolfine in S. brevicaulis. These additive and synergistic interactions indicate that combined therapy with topical amorolfine and oral terbinafine is justified. Sublimation of amorolfine (and terbinafine) may enhance the penetration in and through the nail plate, and support treatment efficacy. These invitro results support the notion that combining oral terbinafine and topical amorolfine is beneficial to patients with onychomycosis, particularly if the pathogen is a non-dermatophyte fungus such as S. brevicaulis.

In vitro antifungal activity of polymeric nanoparticles loaded with Euphorbia tirucalli extract.

The therapeutic potential of medicinal plants is known as an alternative in treatment of human affections; in effect, the conventional application of these medicinal sources has several limitations like low bioavailability, solubility and stability, which affect its pharmacological efficacy. In recent decades, extraordinary advances have been made in new drug delivery systems using nanocarriers. This work consisted in determining the in vitro antifungal activity of the methanolic extract of Euphorbia tirucalli formulated in polymeric nanoparticles. The antifungal activity was determined by the microdilution method in 96-well microplates, applying nanoparticles loaded with plant extract (NP-Ext) obtained by nanoprecipitation on clinical isolates of Trichophyton rubrum and T. interdigitalis. Regarding the nanoparticles, the lots used did not present significant differences in their physicochemical characteristics, with a size of 91.885 ± 1.621nm, polydispersity index of 0.152 ± 0.025 and Z-potential of -6.047 ± 0.987. The quantification of the extract in the polymeric matrix was determined by infrared spectroscopy (FTIR), where an efficiency and encapsulation percentage of 22.15 ± 0.82 and 2.95 ± 0.11, respectively, were obtained. The in vitro antifungal activity of the crude and formulated extract was obtained calculating the Minimum Inhibitory Concentration (MIC) of each one; a MIC of 125 µg/mL was obtained against T. rubrum and T. interdigitalis with the crude extract, while a MIC value of 55.55 and 0.1 µg/mL was obtained with NP-Ext, respectively, against these same. Conclusions: biological activity is closely linked to the phytochemical profile of the extract; while the improvement of said potential with the NP-Ext with the dosage form was directly related to the physicochemical characteristics of the nanocarrier.

Transcriptional analysis of Metalloprotease, Metallocarboxypeptidase, Isocitrate lyase, Citrate synthase, Malate synthase and Dipeptidylpeptidase V of Trichophyton rubrum isolates from dermatophytosis patients.

Introduction: Dermatophytes are pathogenic fungi that cause cutaneousinfection of human and animal and grow exclusively on the stratumcorneum, nail and hair. Methods: The present study was conducted on 160 samples from clinicallydiagnosed onycomycosis patients, further subjected to culture from nailsamples of patients attending dermatology OPD of a tertiary care hospital,Delhi from January 2016 to December 2018 Results: In a soy protein culture, a substantial proteolytic activity was seen, which was secreted by Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. This proteolytic activity was 55–75% inhibited by O-phenanthroline, which authenticate that metalloproteases were secreted by all three species. A consensus probe was constructed on previously characterized genes, which encodes metalloproteases (MEP) of the M36 fungalysin family in Aspergillus fumigatus, Aspergillus oryzae and M. canis. From genomic libraries of T. rubrum, T. mentagrophytes and M. canis, T. rubrum & T. Mentagrophytesa five-member MEP family was isolated and also secretes MCP A & MCPB, MEP 3, MEP4, IL, CS, MS AND DPP V of the M14 family accordingto the MEROPS proteolytic enzyme database. Conclusion: This study shows that Metalloprotease, Metallocarboxypeptidase, Isocitrate Lyase, Citrate Synthase, Malate Synthase and Dipeptidylpeptidase V are expressed in Trichophyton Rubrum Isolates from Dermatophytosis Patients.

In vitro Azole Susceptibility Testing of Japanese Isolates of Itraconazole-Resistant Dermatophytes.

We conducted antifungal susceptibility testing on itraconazole (ITCZ)-resistant isolates of Trichophyton interdigitale and Trichophyton rubrum collected from Japanese patients in 2021 and 2022. The aim of the present study was to determine the most effective drug against ITCZ-resistant strains of dermatophytes. In all isolates, the minimum inhibitory concentrations (MICs) were > 32 mg/l for ITCZ, < 0.03 to 0.5 mg/l for ravuconazole (RVCZ), and < 0.03 mg/l for efinaconazole (EFCZ), luliconazole (LUCZ), and terbinafine (TRBF). Thus, in tinea unguium cases with ITCZ-resistant strains, treatment should be switched to TRBF or other azoles with a stronger antifungal efficacy, such as EFCZ, LUCZ, or RVCZ, and treatment must continue until the infectious organisms are completely eliminated.

Combinatorial effect of fluconazole, itraconazole, and terbinafine with different culture extracts of Candida parapsilosis and Trichophyton spp. against Trichophyton rubrum.

Onychomycosis is a nail infection caused by dermatophytes, non-dermatophyte fungi, and yeasts, especially Candida species. The present study evaluated the combinatorial effect of different cultured extracts of Candida parapsilosis and Trichophyton mentagrophytes and Trichophyton rubrum with fluconazole, itraconazole, and terbinafine against clinical isolates of Trichophyton rubrum. In addition, investigation of the action of the extracts on the wall or membrane was performed. Pure and mixed cultures of Candida parapsilosis and dermatophytes were filtered through a 0.2-μm membrane and submitted to liquid-liquid extraction using ethyl acetate. After a checkerboard, trial with drugs was performed to evaluate the synergistic interaction with the extract. The results obtained for the minimum inhibitory concentration (MIC) of extracts against the T. rubrum strain in isolation were 500-8000 μg/mL. The MIC range for fluconazole, itraconazole, and terbinafine were 2-32 μg/mL, 0.25-0.5 μg/mL, 0.03-64 μg/mL, respectively. However, when the extract was combined with drugs, the MIC values decreased: extracts 1.9-1000 μg/mL, fluconazole 0.25-4, itraconazole 0.03-0.06 μg/mL, and terbinafine 0.001-0.02 μg/mL. The MIC values of the extracts in the Roswell Park Memorial Institute 1640 medium (RPMI) supplemented with sorbitol did not change, suggesting any action on the cell wall. However, in the presence of RPMI supplemented with ergosterol, MIC values of the extracts increased by up to 2×, indicating action on the fungal cell membrane. A synergistic action was observed between products and drugs, detecting a decrease in MIC values. There is potential and a new therapeutic perspective for fungal control.

To Study The Antifungal Susceptibility Pattern, Molecular Profiling And Its Sequencing Analysis Of Trichophyton Rubrum Isolated From Human Sample: A Cross Sectional Case Study From Uttar Pradesh, India

Introduction: There has been a sudden, unexplained surge in dermatophytoses in India since past 5 years. T. rubrum being the most common etiological agent isolated from clinically diagnosed lesions of cutaneus dermatophytosis. It has a significantly higher capacity of transmission than other anthropophilic dermatophytes. Aim and Objective: To study the Antifungal Susceptibility Pattern , Molecular Profiling and its Sequencing Analysis of Trichophyton rubrum isolated from human Sample. Material and Methods: This was a a prospective study carried out in the Department of Microbiology at Rama Medical College Hospital and Research Centre Mandhana, Kanpur for a period of 1 year i.e, March 2017 to March 2018. A total of 375 clinically suspected cases of superficial fungal infection were collected with proper informed consent. Dermatophytosis was confirmed in cases by direct microscopy and culture methods. Antifungal drug susceptibility profiles, MIC of isolates were 50 90 determined using the broth microdilution method according to the CLSI guidelines. The DNA isolation was done using the Qiagen DNA extraction kit followed by the PCR. The sequencing methods were used for the genetic analysis of virulence gene for the confirmation. Results: Out of the total of 375 clinically suspected cases of superficial fungal infection 31 isolates of Trichophyton rubrum were studied. The ratio of Male were more 27 (87%) as compared to the Female 4 (12.9% ). The maximum number of isolates was found in the Skin 28 (90.3%) followed by the Nail 3 (9.6%). The ITZ was most sensitive toward T. rubrum than KTZ, TBF, and FCZ. The molecular characterization for the detection of 18s gene was performed which was confirmed by the Sequencing Analysis. Conclusion: Antifungal resistance against T. rubrum is increasing over the last few decades due to genetic diversity in the fungal genome. The molecular identification is a gold standard technique for the identification of T. rubrum allowing important factor in providing fungal infection prevention and treatment approaches.

Molecular study and antifungal susceptibility profile of Trichophyton rubrum and Trichophyton mentagrophytes strains isolated from lesions of humans and cattle.

Monitoring of contagious diseases is important to advance our knowledge of their epidemiology and to enable more impressive investigation and prevention efforts. This study aimed to examine antifungal drug susceptibility and molecular analysis of clinical isolates of Trichophyton rubrum and Trichophyton mentagrophytes in humans and cattle. A total of 400 patients and 500 cattle were evaluated in this study. Dermatophytosis was confirmed in cases by direct microscopy and culture methods. Antifungal drug susceptibility profiles, MIC50, and MIC90 of isolates were determined using the broth microdilution method. Multiplex-PCR, RAPD PCR, and sequencing methods were used for the genetic analysis of virulence genes and the ITS1 and ITS2 regions, respectively. A total of 175 patients and 120 cattle were diagnosed with dermatophytosis. Dermatophytes showed a remarkable rate (30%) of terbinafine resistance. T. mentagrophytes showed lower susceptibility than T. rubrum (MIC50 =16 μg/mL). Strains harboring Mep1, Mep2, and Mep4 genes had the highest frequency among all genotypes. A RAPD-PCR dendrogram divided T. mentagrophytes and T. rubrum strains into three and six groups, respectively. A notable rate of resistance to terbinafine in isolated dermatophytes was reported in this study. Examination of RAPD-PCR results showed that T. rubrum strains had higher genetic diversity than T. mentagrophytes. Genetic monitoring of dermatophytes must be considered an important factor in providing fungal infection prevention and treatment approaches.

In vitro antifungal activity of Candida culture extracts against Trichophyton rubrum and Trichophyton mentagrophytes.

Onychomycoses are nail infections caused primarily by dermatophytes fungi, yeasts, and other filamentous fungi, characterized by persistent infections, prolonged therapy, and high recurrence rates. In clinical practice, some of these occurrences present two or more microorganisms, and the interactions among them can change the chemical environment mediated by small diffusible molecules, producing a competitive niche. The aim of this study was to evaluate the antifungal activity of individual extracts of pure cultures of Candida albicans and C. parapsilosis against dermatophytes. To obtain the fungal extracts, cultures were filtered through a 0.2 μm membrane and submitted to liquid-liquid extraction using ethyl acetate. The Minimal Inhibitory Concentration (MIC) of each extract was evaluated by broth microdilution method and checkerboard assay with fluconazole against clinical isolates of Trichophyton rubrum and T. mentagrophytes. The invertebrate model of Galleria mellonella was used to evaluate the toxicity of the extracts. As results, the extracts of C. albicans and C. parapsilosis showed antifungal activity with MICs between 31,2 – 2000 μg/mL. In association with fluconazole, synergistic effect was detected for all combinations. The extracts presented low toxicity in G. mellonella. In the future, isolation and identification of the extract compounds may allow new therapeutic approaches in the control of fungal infections.

In vitro MIC comparison of a test itraconazole brand versus 4 brands of itraconazole against clinical isolates of Trichophyton mentagrophytes and Trichophyton rubrum using broth dilution method and disc diffusion method

Dermatophytosis is one of the most common cutaneous fungal infections in India. Despite, quite a few numbers of antifungal agents available for treatment, the resistance to antifungals by dermatophytes is increasing day by day as seen by doctors in clinical practice. Itraconazole has shown good results in the treatment of dermatophytosis at doses of 100 mg once a day for 2 weeks and with 200 mg once a day for 7 days. However, there are multiple itraconazole brands available in India. In this study, five different itraconazole brands were evaluated in vitro for MIC values against clinical isolates of Trichophyton mentagrophytes and Trichophyton rubrum by two different methods i.e. broth dilution and disc diffusion tests. It was found that the Reference brand of itraconazole and the test brand of Itraconazole showed least MIC values as compared to other available brands.

Molecular Docking-Guided Ungual Drug-Delivery Design for Amelioration of Onychomycosis.

The present work envisaged an adherent luliconazole-loaded bilayer nail lacquer (BNL) with significant transungual activity. The locally applied sustained-release BNL formulation was designed for an improved retention, payload, and final dermatokinetic disposition. A primary step in the fabrication of a BNL included overcoming of physical barriers like α-keratin (also α-keratin), a protein present in human nails, and then allowing the drug molecule to permeate at the site of action. Although luliconazole is an established antifungal agent, has limited clinical exploitation for its use in treating onychomycosis. An in silico study elucidating its interaction with lanosterol-14-α demethylase, an enzyme which is the key region of drug action mechanism, was highly supportive of its imminent clinical potential. Optimization of prepared BNL formulations via response surface modeling (Box–Behnken Design-Expert 10.0.6) logically ascertained the effect of selected independent variables and showcased its effect via dependent responses. Surface morphology of the prepared BNL films was well corroborated for the presence of two distinct polymeric layers through scanning electron microscopy imaging. Nail permeation studies revealed a cumulative drug release of 71.25 ± 0.11% through bovine hooves up to 24 h. Luliconazole while exposing antifungal activity against clinical isolates of Trichophyton rubrum in agar cup-plate method disclosed a 38 mm diameter zone of inhibition. Further, the optimized BNL exhibited a bioadhesive force of 1.9 ± 0.11 N, which assured its retention on the nail surface for prolonged duration of time. In Conclusion, it is deduced that the conventional treatment modalities for onychomycosis require circumvention of certain pharmacotechnical caveats. Therefore, in the present study, a multipronged BNL system was proposed, which negates the need of frequent drug application, improvises cosmetic appearance, yields fruitful therapeutic outcomes, and has a clinical supremacy over the available therapeutics.

In vitro susceptibility of seven antifungal agents against dermatophytes isolated in İstanbul

Background/aim: Dermatophytes are the causative agents of dermatophytosis, which is a common disease worldwide that affects the hair, skin, and nails. Dermatophytes comprise more than 40 species in 3 genera: Microsporum, Trichophyton, and Epidermaphyton. In this study, we aimed to determine the effectiveness of seven antifungal agents: amphotericin B, terbinafine, itraconazole, voriconazole, ketoconazole, miconazole, and fluconazole. Materials and methods: A sensitivity study was performed using a microdilution method in accordance with the CLSI M38-A2 standards using isolates of Trichophyton rubrum (n = 55), Microsporum canis (n = 9), and Trichophyton interdigitale (n = 2), which were identified by sequencing the internal transcribed spacer region of the rDNA. Results: According to the results of antifungal sensitivity tests, the geometric mean (GM) minimum inhibitory concentration (MIC) against T. rubrum was 0.10 μg/mL for ketoconazole, 0.20 μg/mL for itraconazole, 0.07 μg/mL for miconazole, 0.48 μg/mL for fluconazole, 2.27 μg/mL for amphotericin B, 0.06 μg/mL for voriconazole, and 0.06 μg/mL for terbinafine. Conclusion: The most effective antifungal drugs were voriconazole and terbinafine, both of which had a GM MIC of 0.06 μg/mL.

Study genetic diversity between trichophyton rubrum isolates using ISSR and RAPD markers

The present study included detection of genetic variability, and identification of genetic relationship and finding a fingerprint of the ten clinical isolates related to Trichophyton rubrum using RAPD and ISSR markers. The experiment was carried out and the results performed using six primer of the RAPD markers these primers showed 239 amplified band , out of these band 90 of them was considered as a main band, and 149 was Polymorphic band , the largest number of bands was 30 in the isolate TR6 and less number of bands in the isolate TR7. The results clear the value of genetic diversity based on RAPD analysis the lowest value of genetic diversity (0.13005) between the isolate TR3 and TR9while the highest genetic diversity (0.55941) was between the isolates TR5 and TR8. The analysis of the relationship shows that there are three main groups: the first group include isolate TR8, while the second group included three isolates are isolate TR2, isolate TR10 and isolates TR3, The third group included three subgroups included the first isolate TR1,isolates TR4 and second isolate TR3 and isolate TR9 and the third included the isolate TR5 and isolates TR6.The results of ISSR experiments: the use six a primers in the of the ISSR showed 192 bands, in the isolate of Trichophyton rubrum, two of these primers showed monomorphic bands (in number and location) and six primers showed monomorphic and polymorphic bands, while one showed only polymorphic bands among Trichophyton rubrum isolates. And the largest number of bands was 24 in the TR5 and less number of bands 16 in isolate TR3 and isolate TR8 were finding DNA fingerprint to isolate TR1, isolate TR5, and isolate TR3. The ISSR markers showed lowest genetic polymorphism was (0.05561) between the isolates TR2 and isolate TR7 and the largest genetic distance was (0.40501) between the isolate TR4 and isolate TR8. The analysis of the relationship of genetic showed that there are three groups key first included isolates and only one isolate TR8 and the second involving isolates TR2 and isolate TR7, isolate TR4 and isolate TR3, while the group included three sub-groups, the first included isolate TR1 and isolate TR6, and the second involving isolate TR5, isolate TR10 and isolate TR9. The results confirmed the efficiency markers of the RAPD and ISSR in contrast, find a genetic fingerprint to the isolates of Trichophyton rubrum, and these markers differ in mechanical detection contrast, and coverage is Genome. Therefore Complementary to each other, although the ISSR markers were more efficient in terms of the number of binding sites in each type of the isolates of Trichophyton rubrum and so can the initiator of the discovery of a much larger area from Genome.

Bioethanol Production from Fungal Treated Rice Husks Fermented with Bakers Saccharomyces cerevisiae and Yeast Isolates from Palm Wine

This research was necessitated by the quest to create a useful product from wastes (bio-ethanol from rice husk). This if successful will serve two purposes; first, help reduce wastes in the environment and to create wealth from waste. Separated isolates of Trichophyton soudanense, Trichophyton rubrum, Trichophyton mentagrophyte, Aspergillus oryzae, Aspergillus niger and Aspergillus fumigatus were obtained from husks of processed rice undergoing decomposition for more than 8 months. Husks of freshly processed rice were pretreated by autoclave boiling for 20 minutes at a temperature of 121°C after mixing in Mandle’s media. The experimental test samples consisted of co-culture combinations and monocultures individually inoculated into various measured heat treated husks; additional control groups were also made. Non-reducing sugar, reducing sugar and total sugar were assayed at the seventh day following hydrolysis. The resulting filtrates of the various husks (treated and control experimental units) were subjected to 7 days fermentation with yeasts from palm wine as well as bakers’ yeasts. Values of the result indicated highest trends in the following treatments: T. mentagrophyte treated husks with soluble reducing sugar value of 2.66 ± 0.14 g/L, A. fumigatus treated husk with soluble non reducing sugar value of 18.08 ± 2.61%, co-culture of T. soudanense and T. rubrum treated husks gave total sugar value of 20.53 ± 2.73%. Fermented A. oryzae treated husk filtrate inoculated with palmwine yeasts had optimal bio-ethanol yield (120.82 ± 0.39 g/L) followed by A. oryzae and T. soudanense treated husks fermented with bakers yeasts with 60.60 ± 0.10 g/L bioethanol. Recognizable yields of bioethanol from palm wine yeast fermented husk as well as sugar from other treated husks were obtained.

Quantitative and structural analyses of the in vitro and ex vivo biofilm-forming ability of dermatophytes.

The aim of this study was to evaluate the in vitro and ex vivo biofilm-forming ability of dermatophytes on a nail fragment. Initially, four isolates of Trichophyton rubrum, six of Trichophyton tonsurans, three of Trichophyton mentagrophytes, ten of Microsporum canis and three of Microsporum gypseum were tested for production biomass by crystal violet assay. Then, one strain per species presenting the best biofilm production was chosen for further studies by optical microscopy (Congo red staining), confocal laser scanning (LIVE/DEAD staining) and scanning electron (secondary electron) microscopy. Biomass quantification by crystal violet assay, optical microscope images of Congo red staining, confocal microscope and scanning electron microscope images revealed that all species studied are able to form biofilms both in vitro and ex vivo, with variable density and architecture. M. gypseum, T. rubrum and T. tonsurans produced robust biofilms, with abundant matrix and biomass, while M. canis produced the weakest biofilms compared to other species. This study sheds light on biofilms of different dermatophyte species, which will contribute to a better understanding of the pathophysiology of dermatophytosis. Further studies of this type are necessary to investigate the processes involved in the formation and composition of dermatophyte biofilms.

Isolation and Molecular Identification of Trichophyton rubrum var.raubitschekii from the Infant Groin.

Five clinical isolates of Trichophyton rubrum isolated from patients withdermatophytosis in the Dermatology unit of the General Hospital in Kalar distric/Sulaimaniprovince/North region of Iraq. The isolates were identified according to the conventionallaboratory methods and molecular methods, one of these strains was isolated from the groinof an infant with one year old and identifies as Trichophyton rubrum var. raubitschekii. Thisvariety characterized by the production of cottony to a velvety colony. The front of the colonyappeared brown color with brownish pigmentation at the reverse. Slide examination revealednumerous cylindrical shaped microconidia and few longer macroconidia. Molecularidentification was conducted according to the conventional PCR by using set primers ITS1and ITS4 and resulted in PCR product about 690bp. PCR-RFLP by using BstN1 digestionenzyme revealed three pattern bands 380,180,100bp. While sequencing of the ITS regionrevealed high similarity with that of a number of uncultured endocytic fungus clone.

Investigation of the antifungal potential of linalool against clinical isolates of fluconazole resistant Trichophyton rubrum

The aim of this study was to investigate the activity of the monoterpene linalool against clinical isolates of Trichophyton rubrum. Initially, a sensitivity assay for commercial antifungals with solid disks in diffusion medium was performed. Minimum inhibitory concentration (MIC) of linalool and ketoconazole (positive control) were determined by microdilution in RPMI 1640 medium (CLSI M38-A2). We then evaluated the action of linalool and ketoconazole at different concentrations (1/2MIC, MIC and 2×MIC) on mycelial growth (radial mycelial growth), conidia production and conidia germination using a hemacytometer. The effects on cell membrane (release of intracellular material) were also investigated. Finally, changes in fungal morphology as induced by the test drugs were analyzed. Based on the sensitivity tests, the fungal strains showed resistance to 5-fluorocytosine and fluconazole. The linalool MIC values ranged from 256μg/mL to 512μg/mL, whereas ketoconazole showed values of 4μg/mL to 8μg/mL. For the LM 305 strain, the test drugs showed the following MIC values: linalool 256μg/mL and ketoconazole 8μg/mL. The mycelial growth of T.rubrum LM 305 was inhibited by linalool (2×MIC) and ketoconazole (1/2MIC, MIC, 2×MIC), in 7 days of treatment (P<0.05). The test-drugs also inhibited conidial germination and conidiogenesis (P<0.05). Linalool also caused leakage of intracellular material (P<0.05). Finally, we verified the effectiveness of linalool and ketoconazole to induce micro-morphological changes, forming abnormal, wide, short and crooked hyphae. Based on these results, we conclude that linalool presents as an antifungal agent with anti-Trichophyton rubrum potential, an important dermatophytosis agent.

Actividad antifúngica de extractos de biomasa celular de neem sobre aislamientos de dermatofitos

Cell lines of neem (Azadirachta indica A. Juss.) grown in liquid suspension have shown to produce bioactive secondary metabolites, particularly triterpenoids. In consequence, its use as a control of medical microorganisms (like dermatophytes) is proposed. The main goal of this study was to assess the antifungal activity of methanolic extracts from neem cultured cell suspensions on several isolates of Trichophyton mentagrophytes (five isolates), Trichophyton rubrum (five isolates) and Epidermophyton floccosum (three isolates). Neem cell suspension cultures were scaled up, from which a raw methanolic extract was obtained. This extract was fractionated by silica gel column chromatography. The raw methanolic extract and its fractions were used in order to determine the Minimal Inhibitory Concentration (MIC) on the dermatophytes isolates by following M38-A2 broth microdilution method. Antimycotic Terbinafine was used as positive control. The results shown that neem raw cellular biomass extract inhibits the growth of T. mentagrophytes, T. rubrum and E. floccosum in at least 100%. In the evaluations of the separated fractions, it was observed that the low polarity fractions had higher antifungal activity (MIC=109 μg/mL) than the raw extract per se (MIC=2500 μg/mL) and the most polar ones (MIC=7000 μg/mL). The latter suggest that neem cells cultured in liquid suspension produces compounds with antifungal activity, being more active those present in the low polarity fractions.

In Vitro Antifungal Susceptibility Of Griseofulvin, Fluconazole, Itraconazole And Terbinafine Against Clinical Isolates Of Trichophyton Rubrum And Trichophyton Mentagrophytes

Investigations on antifungal drug susceptibility were carried out on 90 clinical isolates of Trichophyton rubrum, and Trichophyton mentagrophytes with four antifungal drugs, namely griseofulvin, fluconazole, itraconazole and terbinafine as suggested by National Committee for Clinical Laboratory Standards (NCCLS) M27–A (1997) document by broth macrodilution method to standardize in vitro antifungal susceptibility testing and to find out the Minimum Inhibitory Concentration (MIC) of the drugs. In this study, terbinafine was found to be the most efficient drug for all isolates. Terbinafine had the lowest MIC range of 0.001 g/ml to 0.09 g/ml and MIC50 was low at 0.005 g/ml and MIC90 was also low at 0.04 g/ml against T.rubrum; and MIC range of 0.001μg/ml to 0.19μg/ml with a MIC50 of 0.01μg/ml and MIC90 at 0.09μg/ml against T.mentagrophytes. Itraconazole showed antifungal activity superior to that of fluconazole, with a MIC range of 0.04g/ml to 1.56g/ml, with MIC50 at 0.19μg/ml and MIC90 at 1.56g/ml against T.rubrum; and MIC range of 0.04μg/ml to 1.56μg/ml, with MIC50 at 0.19μg/ml and MIC90 at 0.78μg/ml against T.mentagrophytes. Griseofulvin appears to be still a potent drug for management of dermatophytoses. Griseofulvin had a MIC range of 0.15g/ml to 5.07 g/ml with MIC50 at1.26 g/ml and MIC90 at 2.53 g/ml against T.rubrum; and MIC range of 0.31μg/ml to 5.07μg/ml with MIC50 at 1.26μg/ml and MIC90 at 2.53μg/ml against T.mentagrophytes. Fluconazole showed a high MIC range of 0.19 g/ml to 50 g/ml and MIC50 was high at 1.56g/ml and MIC90 was also high at 12.5 g/ml against T.rubrum; and a high MIC range of 0.09μg/ml to 25.0μg/ml, with MIC50 at 1.56μg/ml and MIC90 at 12.5μg/ml towards T.mentagrophytes. The technique was found to be easy to perform and reliable with consistent results.