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The incidence of opportunistic yeast infections in humans has been increasing over recent years. These infections are difficult to treat and diagnose, in part due to the large number and broad diversity of species that can underlie the infection. In addition, resistance to one or several antifungal drugs in infecting strains is increasingly being reported, severely limiting therapeutic options and showcasing the need for rapid detection of the infecting agent and its drug susceptibility profile. Current methods for species and resistance identification lack satisfactory sensitivity and specificity, and often require prior culturing of the infecting agent, which delays diagnosis. Recently developed high-throughput technologies such as next generation sequencing or proteomics are opening completely new avenues for more sensitive, accurate and fast diagnosis of yeast pathogens. These approaches are the focus of intensive research, but translation into the clinics requires overcoming important challenges. In this review, we provide an overview of existing and recently emerged approaches that can be used in the identification of yeast pathogens and their drug resistance profiles. Throughout the text we highlight the advantages and disadvantages of each methodology and discuss the most promising developments in their path from bench to bedside., The authors discuss the current status of the use of high-throughput (-omics) technologies on the diagnostics of yeast infections.

Staphylococcal pathogens of chronic relapsing infections, such as cystic pneumosclerosis and osteomyelitis are characterized by atypical morphology of the colonies (atypical variants of staphylococci) and present a subpopulation in clinically significant staphylococci. Since the loss of some phenotypic characteristics important for the genus Staphylococcus due to mutations, identification of such staphylococcal variants is difficult and sometimes impossible. An algorithm of identification of atypical variants of S.aureus (SSCVs) was developed. The advantages of the molecular methods and in particular the tRNA-PCR analysis, as well as the use of the diagnostic preparation Diastaph for correct identification of Staphylococcus atypical variants were shown.

AIM: This 3-month double-blind randomized placebo-controlled study evaluated the clinical and microbial effects of an essential oil mouth rinse used as an adjunct to mechanical plaque control by patients in supportive periodontal care. MATERIAL AND METHODS: Fifty patients were randomly allocated to an essential oil group (Listerine(®) Coolmint; Johnson & Johnson, New Brunswick, NJ, USA) or placebo group to rinse twice per day as an adjunct to mechanical plaque control. At baseline and after 3 months, plaque index (PI), gingivitis index (GI), probing pocket depth, bleeding on probing (BoP) and clinical attachment level were registered. Subgingival plaque samples were collected for the detection and quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Micromonas micros, Prevotella intermedia, Fusobacterium genus and Streptococcus mutans by means of real-time PCR (qPCR). Patient's compliance, satisfaction and side effects were registered. RESULTS: Twenty-three patients in the essential oil group (mean age: 57) and 21 in the placebo group (mean age: 55) with acceptable oral hygiene at intake (mean PI CONCLUSION: Patients in supportive periodontal care who are fairly compliant with oral hygiene may not benefit from additional mouth rinsing using an essential oil solution.

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We describe the small-scale, laboratory-based, production and quality control of a cocktail, consisting of exclusively lytic bacteriophages, designed for the treatment of Pseudomonas aeruginosa and Staphylococcus aureus infections in burn wound patients. Based on successive selection rounds three bacteriophages were retained from an initial pool of 82 P. aeruginosa and 8 S. aureus bacteriophages, specific for prevalent P. aeruginosa and S. aureus strains in the Burn Centre of the Queen Astrid Military Hospital in Brussels, Belgium. This cocktail, consisting of P. aeruginosa phages 14/1 (Myoviridae) and PNM (Podoviridae) and S. aureus phage ISP (Myoviridae) was produced and purified of endotoxin. Quality control included Stability (shelf life), determination of pyrogenicity, sterility and cytotoxicity, confirmation of the absence of temperate bacteriophages and transmission electron microscopy-based confirmation of the presence of the expected virion morphologic particles as well as of their specific interaction with the target bacteria. Bacteriophage genome and proteome analysis confirmed the lytic nature of the bacteriophages, the absence of toxin-coding genes and showed that the selected phages 14/1, PNM and ISP are close relatives of respectively F8, phiKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.