A rapid two-step DNA extraction method and a multiplex PCR for the detection of dermatophytes in general andTrichophyton rubrumspecifically were developed and evaluated with DNA extracted from pure cultures and from clinically diseased nails. DNA from the following dermatophytes was used:Epidermophyton floccosum,Microsporum audouinii,Microsporum canis,Microsporum gypseum,Microsporum nanum,Trichophyton mentagrophytes,Trichophyton rubrum,Trichophyton schoenleinii,Trichophyton soudanense,Trichophyton terrestre,Trichophyton tonsurans,Trichophyton verrucosum, andTrichophyton violaceum. Human DNA and DNA from the following nondermatophyte fungi were included as controls:Alternaria,Aspergillus niger,Candida albicans,Candida glabrata,Candida krusei,Malassezia furfur,Saccharomyces cerevisiae, andScopulariopsis brevicaulis.A total of 118 nail samples received for routine microscopy and culture for dermatophytes were subsequently tested by the two PCRs separately and in a multiplex format. Using DNA extracted from pure cultures and the pan-dermatophyte PCR, theT. rubrum-specific PCR sequentially and in a multiplex format correctly detected all dermatophytes and additionally correctly identifiedT. rubrum.Comparison of the traditional diagnostic evaluation (microscopy and culture) of nail samples with PCR on DNA directly extracted from the nails showed excellent agreement between PCR and microscopy, but the number of samples with dermatophyte species identification was increased considerably from 22.9% to 41.5%, mainly due to the identification ofT. rubrumby PCR in microscopy-positive but culture-negative samples. In conclusion, this 5-hour diagnostic test was shown to increase not only the speed but also the sensitivity of investigation for nail dermatophytosis.

Blastocystis hominis is a common enteric parasite of worldwide distribution. Its pathogenetic potential has not yet been established, although numerous case reports suggest that B. hominis may cause the development of various gastrointestinal symptoms and disorders. The detection of the parasite in stool specimens is conventionally done by microscopy of direct smears, fecal concentrates, or permanently stained smears; however, morphology-based diagnosis is problematic. The aim of this study was to develop and evaluate a polymerase chain reaction (PCR) technique for the direct detection of B. hominis in human stool samples. Primers were based on small subunit ribosomal DNA and able to detect > or =32 parasites/200 mg stool artificially spiked with cultured B. hominis. In the evaluation of 43 clinical specimens, the PCR was tested against the formol ethyl acetate concentration technique (FECT) and a culture technique, proving 100% test specificity and a significantly higher sensitivity than the FECT. The PCR method is recommended for screening clinical specimens for B. hominis infection and for use in prevalence studies.

Fungal infections are rising all over the world every year. There are only five medical compound classes for treatment: triazoles, echinocandins, polyenes, flucytosine and allylamine. Currently, echinocandins are the most important compounds, because of their wide activity spectrum and much lower sides effects that may occur during therapy with other drugs. Echinocandins are secondary metabolites of fungi, which can inhibit the biosynthesis of β-(1,3)-D-glucan. These compounds have fungicidal and fungistatic activity depending on different genera of fungi, against which they are used. Echinocandin resistance is rare—the major cause of resistance is mutations in the gene encoding the β-(1,3)-D-glucan synthase enzyme. In this review of the literature we have summarized the characteristics of echinocandins, the mechanism of their antifungal activity with pharmacokinetics and pharmacodynamics, and the resistance issue.

The factors enabling Candida spp. infections are secretion of hydrolytic enzymes, adherence to surfaces, biofilm formation or morphological transition, and fitness attributes. The aim of this study was to investigate the correlation between known extracellular virulence factors and survival of Galleria mellonella larvae infected with clinical Candida. The 25 isolates were tested and the activity of proteinases among 24/24, phospholipases among 7/22, esterases among 14/23, hemolysins among 18/24, and biofilm formation ability among 18/25 isolates was confirmed. Pathogenicity investigation using G. mellonella larvae as host model demonstrated that C. albicans isolates and C. glabrata isolate were the most virulent and C. krusei isolates were avirulent. C. parapsilosis virulence was identified as varied, C. inconspicua were moderately virulent, and one C. palmioleophila isolate was of low virulence and the remaining isolates of this species were moderately virulent. According to our study, virulence of Candida isolates is related to the expression of proteases, hemolysins, and esterases.

The physical/chemical abatement of gas pollutants creates many technical problems, is costly and entails significant environmental impacts. Biological purification of off-gases is a cheap and ecologically safe way of neutralization of gas pollutants. Despite the recent advances, the main technological challenge nowadays is the purification of volatile organic compounds (VOCs) of hydrophobic character due to their low solubility in water. Among all known biological methods of air purification, the most cost-effective biodegradation of hydrophobic VOCs is conducted by biotrickling filters. In this context, fungi have gained an increasing interest in this field based on their ability to biodegrade hydrophobic VOCs. In addition, biotrickling filtration using fungi can support a superior hydrophobic VOC abatement when compared to the bacterial biofilters. This paper aims at reviewing the latest research results concerning biocatalytic activity of fungi and evaluating the possibilities of their practical application in biofiltration systems to remove hydrophobic VOCs.

We recently reported the development of a 5-hour multiplex PCR test for the detection of tinea unguium and the optimization of this test by the inclus