Mycopathologia et mycologia applicata

Mouse model is an appropriate tool for pathogenic determination and study of host defenses during the fungal infection. Here, we established a mouse model of candidiasis with concurrent oral and vaginal mucosal infection. Two C. albicans strains sourced from clinical candidemia (SC5314) and mucosal infection (ATCC62342) were tested in ICR mice. The different combinational panels covering estrogen and immunosuppressive agents, cortisone, prednisolone and cyclophosphamide were used for concurrent oral and vaginal candidiasis establishment. Prednisolone in combination with estrogen proved an optimal mode for concurrent mucosal infection establishment. The model maintained for 1 week with fungal burden reached at least 105 cfu/g of tissue. This mouse model was evaluated by in vivo pharmacodynamics of fluconazole and host mucosal immunity of IL-17 and IL-23. Mice infected by SC5314 were cured by fluconazole. An increase in IL-23 in both oral and vaginal homogenates was observed after infection, while IL-17 only had a prominent elevation in oral tissue. This model could properly mimic complicated clinical conditions and provides a valuable means for antifungal assay in vivo and may also provide a useful method for the evaluation of host–fungal interactions.

In this paper the author has described seventeen new Deuteromycetes from Jabalpur (M.P.) India. These are: Phyllosticta careyae Sahni onCareya arborea Roxb.,Phyllosticta anogeissi Sahni onAnogeissus latifolia Wall.,Phomopsis dalbergiae Sahni onDalbergia sissoo Roxb.,Phomopsis yuccae Sahni onYucca aloifolia L.,Phomopsis dracaenae Sahni onDracaena brachystachys Hook.,Phomopsis buteae Sahni onButea monosperma (Lam.)Kunze,Cytospora grevilleae Sahni onGrevillea robusta A. Cunn.,Coniothyrium sarcinellae Sahni onSarcinella palawanensis (Syd.)Sahni parasitising leaves ofCalastrus paniculatus Willd.,Coniothyrium dioscoreae Sahni on fruits ofDioscorea sp.,Amerodiscosiella indica Sahni onIxora parviflora Vahl.,Ascochyta nyctanthis Sahni onNyctanthes arbor-tristis L.,Pseudodiplodia buteae Sahni onButea monosperma (Lam.)Kunze,Pseudodiplodia oreodoxae Sahni onOreodoxa oleracea Mart.,Phaeoseptoria bougainvilleae Sahni onBougainvillea glabra Choisy,Hainesia jabalpurensis Sahni onWoodfordia fruticosa (L.)Kurz.,Colletotrichum arjunae Sahni onTerminalia arjuna W. & A., andColletotrichum terminaliae Sahni onTerminalia bellirica (Gaertn.)Roxb.

Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) is widely used in clinical laboratories for routine identification of bacteria and yeasts. However, methodological difficulties are still apparent when applied to filamentous fungi. The liquid cultivation method recommended by Bruker Daltonics GmbH for identification of filamentous fungi by MALDI-TOF MS is labour intensive and time-consuming. In this study, growth of Aspergillus species on different (porous) surfaces was investigated with the aim to develop a more reliable, quicker and less laborious identification method using MALDI-TOF MS. Mycelial growth without sporulation mimicking liquid cultivation and reliable MALDI-TOF MS spectra were obtained when A. fumigatus strains were grown on and in between a polycarbonate membrane filter on Sabouraud dextrose agar. A database of in-house reference spectra was created by growing Aspergillus reference strains (mainly focusing on sections Fumigati and Flavi) under these selected conditions. A test set of 50 molecularly identified strains grown under different conditions was used to select the best growth condition for identification and to perform an initial validation of the in-house database. Based on these results, the cultivation method on top of a polycarbonate filter proved to be most successful for species identification. This method was therefore selected for the identification of two sets of clinical isolates that mainly consisted of Aspergilli (100 strains originating from Indonesia, 70 isolates from Qatar). The results showed that this cultivation method is reliable for identification of clinically relevant Aspergillus species, with 67% and 76% correct identification of strains from Indonesia and Qatar, respectively. In conclusion, cultivation of Aspergilli on top of a polycarbonate filter showed improved results compared to the liquid cultivation protocol recommended by Bruker in terms of percentage of correct identification, ease of MSP creation, time consumption, cost and labour intensity. This method can be reliably applied for identification of clinically important Aspergilli and has potential for identification of other filamentous fungi.

Phylogenetic studies highlight Candida africana as an atypical variant within Candida albicans species complex which is dominantly recovered from vaginal specimens. This study aimed to characterize C. africana isolates from patients with vulvovaginal candidiasis (VVC) by molecular methods and in vitro susceptibilities. One hundred and fifty-six (48.44 %) Candida strains were collected from 322 patients diagnosed with VVC. Of these, 114 (73.07 %) were germ tube positive and presented green color on the chromogenic medium, thus classified as C. albicans species complex. One hundred and nine (95.61 %) out of 114 isolates were identified as C. albicans, while five (4.38 %) isolates were identical with C. africana based on hwp1 PCR. C. africana appeared to be highly susceptible to the tested antifungals. For all strains of C. africana, fluconazole MIC was 2-log2-dilution steps less active than amphotericin B, which in turn was 2-log2-dilution steps and 3-log2-dilution steps less active than other azoles and echinocandin agents, respectively. In conclusion, among the C. albicans species complex, C. albicans predominantly and C. africana rarely occur in vaginal mucosa. Due to limited information on molecular epidemiology of this novel yeast, more studies using molecular methods are needed to elucidate the inter- and intraspecific genomic variations of C. africana isolates.

The predominance of non-Candida albicans Candida (NCAC) species causing healthcare-associated infections has increased over the last decade pertaining to their ability to form biofilms on medical devices. These biofilm-associated infections are challenging to treat as they are resistant to antifungal agents and evade host-immune response resulting in a high risk of device failure or biomaterial removal. Thus, to minimize the risk of biofilm-associated infections, preventing biofilm formation is the best approach which is mediated by the quorum quenching process. The present study investigated the modulatory effect of 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF) on NCAC biofilm formation and also assessed the effect of the DMHF-coated catheters on biofilm formation of NCAC. The NCAC isolates studied were Candida tropicalis, Candida glabrata and Candida krusei isolated from catheter tip, urine and blood, respectively. DMHF at a concentration of 30 µg/mL showed an inhibitory effect against NCAC biofilms at various stages and was statistically significant (p ≤ 0.05) against the various concentrations (50–5 µg/mL) tested and also among the three phases of experiment. The furanone content on coated catheters ranged from 170 to 750 µg and release of furanone from the coated catheter was about 15 µg for 30 days. The effect of DMHF-coated catheters on NCAC biofilm formation was observed by the scanning electron microscopy which revealed the absence of NCAC adherence on DMHF-coated catheters. This study provides a design to develop furanone-coated biomaterials which could be implemented in healthcare settings to reduce medical device-associated infections. The excellent biological performance, combined with their antimicrobial properties, suggests that 2,5-dimethyl-4-hydroxy-3(2H)-furanone could be an effective anti-infective coating for implantable devices.

Addition of the essential oil ofHyssopus officinalis to the culture medium ofAspergillus fumigatus induced alterations in both growth and lipid composition of this mould. Total lipids and sterols were reduced, whereas total phospholipids were increased. There were alterations in the proportions of fatty acids, neutral lipid and phospholipid fractions.

The haploid chromosome number ofHypoxylon cohaerens apparently is 5, based on counts made at meiotic prophase and meiotic and mitotic metaphases. Newly formed ascospores are at first uninucleate, becoming binucleate following mitosis in the ascospore. Subsequently, one of the two nuclei disappears. Maturing ascospores are uninucleate.

Fusarium is a ubiquitous hyalohyphomycete isolated from food, widespread in the environment (plants, soil) and present at all latitudes. Fusarium oxysporum and Fusarium solani are the most frequent pathogenic species, followed by F. moniliforme and F. chlamydosporum. Infections due to this mold may be disseminated or localized. Localized forms include cutaneous and subcutaneous infection, onychomycosis, endophtalmitis, otitis, sinusitis, arthritis, osteomyelitis, and brain abscess. Disseminated forms are those in which two or more noncontiguous sites may be involved. These latter are observed in patients with severe neutropenia. Wounds, digital ulcers, onychomycosis, and paronychia are the typical cutaneous portal of entry. We report a case of primary localized cutaneous infection due to Fusarium in a 29-year-old woman presenting with a nodular lesion, partially ulcerated, asymptomatic on the first finger of the left hand, appeared 4 months earlier. Histological examination showed spongiosis and acanthosis in the stratum corneum, ulceration and inflammation with prevalently mononucleate cells and septate and branched fungal structures in the epidermis and in dermis. The fungus was identified as Fusarium oxysporum by culture of biopsy fragments on Sabouraud dextrose agar with chloramphenicol. The culture was deposited in the culture collection of the mycology section of IHEM, Brussels (IHEM21984 col no. 125). The patient had normal immune status and was successfully treated with surgical excision. Recovery was confirmed at follow-up 8 months later.

Eight species ofCephalosporium from man are revised, mostly on original strains.Cryptomyces pleomorpha Gruner is a typicalCephalosporium acremonium; in additionCryptomyces is a valid Phacidiaceous genus described more than a century ago. Of two human strains of the same species, one has been fully pathogenic for laboratory animals. Allantospora violacea Ambr.,A. onychophila Vuill. and an unnamed strain were revised. All are to be referred toCephalosporium (Allantospora)onychophilum (Vuill.) Coudert. Two strains ofAcremonium kiliense are typicalC. acremonium. C. serrae Maffei is a good species, but probably indistinguishable from the saprophyticC. subverticillatum Schulz et Sacc., reisolated in Italy in the year 1934. C. spinosum Negroni — probably indistinguishable fromC. cordoniformis Barbosa, both South American species — is a coremioidal form ofC. acremonium. C. rubrobrunneum (?) Benedek is a simpleC. acremonium, as well asC. stuehmeri Schmidt et van Beyma. In addition to the previously named species,C. pseudofermentatum Cif. (if really possess a yeast-like stage) andC. nigrum Kamb. (if this species is pertaining to the genusCephalosporium) are listed.