Direct identification of molds by sequence analysis in fungal chronic rhinosinusitis.

Fungi are widely implicated in chronic rhinosinusitis. Direct microscopic examination (DME) is used to confirm the biological diagnosis of fungal rhinosinusitis (FRS). Diagnostic sensitivity of DME is better than culture, however DME does not allow fungal species identification. In this study, we included 54 sinus samples demonstrating hyphae on DME. Direct sequencing was compared to culture for the identification of the fungal species. Sequence analysis identified fungi in 81.5% of cases while culture was positive in only 31.5%. The most common genus was Aspergillus and the identified species belonged to section Fumigati or to section Flavi. Among other fungi identified by sequence analysis, Schizophyllum commune was present in three samples attesting to the importance of this Basidiomycetes in FRS. Our results clearly demonstrate the superiority of sequencing compared to culture when performed on specimens with hyphal elements at DME, and contributes to the epidemiological knowledge of fungi involved in FRS.

[Biological diagnosis of onychomycoses. Direct examination after simplified technique of PAS staining]. / Diagnostic biologique des onychomycoses. Examen direct après coloration PAS simplifiée.

Confirmation of fungal origin of onychopathy by mycological examination is essential. For that purpose, in parallel to the cultivation of biological samples, achieving a sensitive and informative direct examination of nail fragments and subungual material is primordial. Among the direct examination techniques, and inspired from a technique of reference in histo-pathology (the "periodic acid-Schiff reagent" reaction), the simplified technique of PAS staining according to Hotchkiss and MacManus is the technique of choice. Easy to implement and very sensitive, it can immediately and formally confirm the diagnosis of onychomycosis, mention the type of fungus (yeast, dermatophyte, Hyphomycete opportunistic) and suspect a possible multiple involvement.

Assessment of caspofungin susceptibility of Candida glabrata by the Etest®, CLSI, and EUCAST methods, and detection of FKS1 and FKS2 mutations.

Candida glabrata has emerged as a major pathogen in invasive candidiasis in recent years. Currently, guidelines for invasive candidiasis treatment recommend fluconazole or an echinocandin as the first-line therapy. Nevertheless, the resistance of Candida glabrata to echinocandin is an emerging problem and has been partly associated with mutations in the FKS1 and FKS2 genes. The Etest® is an appropriate method for determining antifungal susceptibility in emergency routine diagnosis. In this work, we evaluated the reliability of the Etest® in comparison with the two reference broth microdilution methods, Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST), to assess the caspofungin resistance of 193 isolates of Candida glabrata. The interpretation of minimum inhibitory concentration (MIC) values was also discussed according to different breakpoints. Moreover, FKS1 and FKS2 mutations were investigated for isolates with high MICs. Our results showed that the MIC50 value was similar to the MIC90 value for each method. The Etest® method showed the lowest MIC values, whereas EUCAST presented the highest. Categorical agreement between the Etest® and CLSI methods was 100 % and 36 % using the breakpoints proposed by Arendrup et al. (Antimicrob Agents Chemother 56(7):3965-3968, 2012) and Pfaller et al. (Int J Antimicrob Agents 38(1):65-69, 2011), respectively. Two isolates showed high MIC values with the three methods and both presented FKS2 mutations. A novel FKS2 mutation was also reported for one isolate. Future epidemiological studies should also evaluate the reliability of the Etest® to detect echinocandin resistance, as it remains a routine method.

Evaluation of a new test for candidiasis diagnosis in elderly people.

OBJECTIVE: The purpose of this study was to evaluate a new test for candidiasis diagnosis (fungi-dent color test) in an institutionalised elderly population. BACKGROUND: Yeasts normally exist in the human body flora and exploit a situation of weakness in the host. Epidemiologic analyses of yeast infection in the oral cavity showed that the most frequently involved species were Candida albicans. MATERIALS AND METHODS: This study included 73 persons in the short-term and long-term geriatric department of the Montpellier hospital. Oral examinations were performed and the presence of Candida was evaluated: the fungi-dent Color diagnostic method was compared both to the classical culture analysis method, considered as the gold standard, and to the visual clinical diagnostic. RESULTS: The fungi test yielded a sensitivity value of 64.6% (CI: 53.6% - 75.6%) and a specificity of 96.0% (CI: 92.0% - 100%), whereas the diagnostic method through clinical examination disclosed only 22.5% candidiasis. The area under the ROC curve was equal to 0.81. The mean pH value was significantly higher when candidiasis was present (6.15 vs 5.78, p = 0.007). CONCLUSION: Statistical analyses showed that the Fungi-dent-color test was eligible to be validated for community use, with better characteristics than a routine clinical diagnosis and with a reasonably rapid and reliable diagnostic outcome.

Seven imported histoplasmosis cases due to Histoplasma capsulatum var. capsulatum: From few weeks to more than three decades asymptomatic period.

Histoplasma capsulatum is a nonendemic fungus in Europe. Epidemiological, clinical biological features and follow-up of seven imported cases in France were reported. These cases underlined the difficulties of such diagnosis in nonendemic areas due to greatly variable asymptomatic period, and lack of specific clinical signs. Thus, anamnesis should be carefully analysed; mucocutaneous signs and immunosuppression should be systematically investigated. Biological diagnosis requires multiple sites sampling and long-term conservation of culture medium.

Antifungal susceptibility of 205 Candida spp. isolated primarily during invasive Candidiasis and comparison of the Vitek 2 system with the CLSI broth microdilution and Etest methods.

Infections due to Candida spp. are frequent, particularly in immunocompromised and intensive care unit patients. Antifungal susceptibility tests are now required to optimize antifungal treatment given the emergence of acquired antifungal resistance in some Candida species. An antifungal susceptibility automated method, the Vitek 2 system (VK2), was evaluated. VK2 was compared to the CLSI broth microdilution reference method and the Etest procedure. For this purpose, 205 clinical isolates of Candida spp., including 11 different species, were tested for fluconazole, voriconazole, and amphotericin B susceptibility. For azoles, essential agreement ranged from 25% to 100%, depending on the method used and the Candida species tested. Categorical agreements for all of the species averaged 92.2% and ranged from 14.3 to 100%, depending on the 24-h or 48-h MIC reading by the Etest and CLSI methods and on the Candida species. Results obtained for Candida albicans showed excellent categorical and essential agreements with the two comparative methods. For Candida glabrata, the essential agreement was high with the CLSI method but low with the Etest method, and several very major errors in interpretation were observed between VK2 and the Etest method for both azoles. Low MICs of fluconazole were obtained for all of the Candida krusei isolates, but the VK2 expert software corrected all of the results obtained to resistant. Amphotericin B results showed MICs of < or = 1 mg/liter for 201 (VK2), 190 (CLSI), and 202 (Etest) isolates. The AST-YS01 Vitek 2 card system (bioMérieux) is a reliable and practical standardized automated antifungal susceptibility test. Nevertheless, more assays are needed to better evaluate C. glabrata fluconazole sensitivity.

Aspergillus flavus mitral valve endocarditis after lung abscess.

A 16-year-old male with bone marrow failure due to chemotherapy for recurrent acute lymphoblastic leukemia developed an abscess in the lower lobe of the left lung draining through a bronchogastric fistula, as well as mitral valve endocarditis with large vegetations. After a course of antifungal therapy, the left lobe was removed and the fistula closed. The mitral valve was then replaced, after a failed attempt at valve repair, by a mechanical, double-leaflet prosthesis. Microscopy of the lung and heart specimens disclosed hyphae. Cultures of both specimens on Sabouraud's medium recovered a fungus, which was identified by culturing on Czapek's medium as Aspergillus flavus. Despite further antifungal therapy, fatal embolism developed. The emboli contained the same A. flavus as the valve and lung specimens. This case confirms the grim prognosis of primary Aspergillus endocarditis in immunocompromised patients, and suggests that delayed surgical treatment and the presence of another focus of Aspergillus infection may increase the risk of death.

Numerical taxonomy of Old World Phlebotominae (Diptera: Psychodidae). 1. Considerations of morphological characters in the genus Phlebotomus Rondani & Berté 1840.

Numerical analyses (correspondence analysis, ascending hierarchical classification, cladistic approach) were applied to the morphological characters of the adults of the genus Phlebotomus Rondani & Berté 1840. They confirm the reliability of the classic classifications, and also redefine the taxonomic and phylogenetic position of certain taxa. Thus, Spelaeophlebotomus Theodor 1948. Idiophlebotomus Quate & Fairchild 1961 and Australophlebotomus Theodor 1948 deserve generic rank. Among the vectors of leishmaniasis, the subgenus Phlebotomus Rondani & Berté 1840 is probably ancient. The results attribute an intermediate taxonomic and phylogenetic position to the taxa Euphlebotomus Theodor 1948 and Anaphlebotomus Theodor 1948, and reveal the probable artificial nature of the latter. The comparatively large numbers of species of subgenera Paraphlebotomus Theodor 1948, Synphlebotomus Theodor 1948 and, above all, Larroussius Nitzulescu 1931 and Adlerius Nitzulescu 1931, suggest that they are relatively recent. The development of adult morphological characters, the validity of their use in taxonomy and proposals for further studies are discussed.

Numerical taxonomy of Old World Phlebotominae (Diptera: Psychodidae). 2. Restatement of classification upon subgeneric morphological characters.

Numerical analyses (correspondence analysis, ascending hierarchical classification, and cladistics) were done with morphological characters of adult phlebotomine sand flies. The resulting classification largely confirms that of classical taxonomy for supra-specific groups from the Old World, though the positions of some groups are adjusted. The taxa Spelaeophlebotomus Theodor 1948, Idiophlebotomus Quate & Fairchild 1961, Australophlebotomus Theodor 1948 and Chinius Leng 1987 are notably distinct from other Old World groups, particularly from the genus Phlebotomus Rondani & Berté 1840. Spelaeomyia Theodor 1948 and, in particular, Parvidens Theodor & Mesghali 1964 are clearly separate from Sergentomyia França & Parrot 1920.

[Parasitic fecal analyses. Prescription, application and interpretation of results]. / Les analyses coprologiques parasitaires. Prescription, mise en oeuvre et interprétation des résultats.

The increasing ease of intercontinental travel, immigration and the multiplication of cases of immune depression are such that European physicians are now frequently required to prescribe stool examinations for parasites. This article defines the laboratory approach to the diagnosis of intestinal parasites: indications for tests, based upon clinical and epidemiological data; methods of prescription and recommendations to patients; techniques used in the laboratory and their cost; minimum answer times; interpretation of results, in particular in the immune depressed.

[Notes on the phlebotomus of Aïr (Niger)]. / Notes sur les phlébotomes de l'Aïr (Niger).

238 phlebotomine sandfly belonging to seven species have been collected in four localities of Niger, in the Aïr mountain range. Phlebotomus bergeroti was the dominant species in animal bait and in collections inside dwellings. Some attention has to be paid to P. alexandri, a recognized vector of kala-azar elsewhere.

Application of numerical taxonomic methods on phlebotominae.

Numerical analyses (factorial analysis, ascending hierarchical classification, and cladistic) were carried out on morphological characters of adult phlebotomine sandflies. Contributions emerge to the taxonomy and phylogeny of the group. Classical taxonomy is largely confirmed for Old World groups, though the positions of some are adjusted.

[Simultaneous presence in dogs of 2 zymodemes of the Leishmania infantum complex]. / Présence simultanée chez le chien de deux zymodèmes du complexe Leishmania infantum.

The concomitant presence of two zymodemes of the leishmania infantum complex. MON-1 and MON-77, is reported in a dog with diffuse leishmaniasis. The zymodemes were present in both the skin and lymph nodes. Possible explanations for the presence of the two zymodemes are discussed.