Showing papers in "Medical Mycology in 2009"

Consensus multi-locus sequence typing scheme for Cryptococcus neoformans and Cryptococcus gattii.

Abstract: This communication describes the consensus multi-locus typing scheme established by the Cryptococcal Working Group I (Genotyping of Cryptococcus neoformans and C. gattii) of the International Society for Human and Animal Mycology (ISHAM) using seven unlinked genetic loci for global strain genotyping. These genetic loci include the housekeeping genes CAP59,GPD1, LAC1, PLB1, SOD1, URA5 and the IGS1 region. Allele and sequence type information are accessible at http://www.mlst.net/ .

Biofilms of non-Candida albicans Candida species: quantification, structure and matrix composition.

Abstract: Most cases of candidiasis have been attributed to C. albicans, but recently, non-Candida albicans Candida (NCAC) species have been identified as common pathogens. The ability of Candida species to form biofilms has important clinical repercussions due to their increased resistance to antifungal therapy and the ability of yeast cells within the biofilms to withstand host immune defenses. Given this clinical importance of the biofilm growth form, the aim of this study was to characterize biofilms produced by three NCAC species, namely C. parapsilosis, C. tropicalis and C. glabrata. The biofilm forming ability of clinical isolates of C. parapsilosis, C. tropicalis and C. glabrata recovered from different sources, was evaluated by crystal violet staining. The structure and morphological characteristics of the biofilms were also assessed by scanning electron microscopy and the biofilm matrix composition analyzed for protein and carbohydrate content. All NCAC species were able to form biofilms although these were less extensive for C. glabrata compared with C. parapsilosis and C. tropicalis. It was evident that C. parapsilosis biofilm production was highly strain dependent, a feature not evident with C. glabrata and C. tropicalis. Scanning electron microscopy revealed structural differences for biofilms with respect to cell morphology and spatial arrangement. Candida parapsilosis biofilm matrices had large amounts of carbohydrate with less protein. Conversely, matrices extracted from C. tropicalis biofilms had low amounts of carbohydrate and protein. Interestingly, C. glabrata biofilm matrix was high in both protein and carbohydrate content. The present work demonstrates that biofilm forming ability, structure and matrix composition are highly species dependent with additional strain variability occurring with C. parapsilosis.

Occurrence and relevance of filamentous fungi in respiratory secretions of patients with cystic fibrosis – a review

Abstract: The colonization of airways by filamentous fungi and the development of respiratory infections require some predisposing factors as encountered in patients with cystic fibrosis (CF). Indeed, the defective mucociliary clearance which characterizes the disease is associated with local immunological disorders. In addition, the prolonged therapy with antibiotics and the use of corticosteroid treatments also facilitate fungal growth. An important fungal biota has been described in respiratory secretions of patients suffering from CF. Aspergillus fumigatus, Scedosporium apiospermum and Aspergillus terreus for filamentous fungi and Candida albicans for yeasts are the main fungal species associated with CF. Although less common, several fungal species including Aspergillus flavus and Aspergillus nidulans may be isolated transiently from CF respiratory secretions, while others such as Exophiala dermatitidis and Scedosporium prolificans may chronically colonize the airways. Moreover, some of them like Penicillium emersonii and Acrophialophora fusispora are encountered in humans almost exclusively in the context of CF. As fungal complications in CF patients are essentially caused by filamentous fungi the present review will not include works related to yeasts. In CF patients, fungi may sometimes be responsible for deterioration of lung function, as occurs in allergic broncho-pulmonary aspergillosis (ABPA) which is the most common fungal disease in this context. Additionally, although the clinical relevance of the fungal airway colonization is still a matter of debate, filamentous fungi may contribute to the local inflammatory response, and therefore to the progressive deterioration of the lung function.

Chromoblastomycosis: an overview of clinical manifestations, diagnosis and treatment

Abstract: Chromoblastomycosis is one of the most frequent infections caused by melanized fungi. It is a subcutaneous fungal infection, usually an occupational related disease, mainly affecting individuals in tropical and temperate regions. Although several species are etiologic agents, Fonsecaea pedrosoi and Cladophialophora carrionii are prevalent in the endemic areas. Chromoblastomycosis lesions are polymorphic and must be differentiated from those associated with many clinical conditions. Diagnosis is confirmed by the observation of muriform cells in tissue and the isolation and the identification of the causal agent in culture. Chromoblastomycosis still is a therapeutic challenge for clinicians due to the recalcitrant nature of the disease, especially in the severe clinical forms. There are three treatment modalities, i.e., physical treatment, chemotherapy and combination therapy but their success is related to the causative agent, the clinical form and severity of the chromoblastomycosis lesions. There is no treatment of choice for this neglected mycosis, but rather several treatment options. Most of the patients can be treated with itraconazole, terbinafine or a combination of both. It is also important to evaluate the patient’s individual tolerance of the drugs and whether the antifungal will be provided for free or purchased, since antifungal therapy must be maintained in long-term regimens. In general, treatment should be guided according to clinical, mycological and histopathological criteria.

Epidemiology and outcome of Scedosporium prolificans infection, a review of 162 cases.

Abstract: Scedosporium prolificans is a truly emerging fungal pathogen. It has only been recognized as a human pathogen for 22 years and has been related with numerous infections in immunocompromised and immunocompetent patients. A search for cases in the literature was performed and a database was constructed. Cases were reviewed in order to analyse the epidemiology and outcome of infection. A total of 162 cases were included. The median age of patients was 45 years (ranging from a few months to 81 years), and 102 (63%) infections were diagnosed in males. Risk factors for scedosporiosis were malignancy, 74/162 (45.7%), cystic fibrosis, 19/172 (11.7%), and solid organ transplantation 14/162 (8.6%). The most common clinical presentations were disseminated infection, 72/162 cases (44.4%), pulmonary mycosis, 47/162 (29%), and bone and joint infections, 17/162 (10.4%). All disseminated infections afflicted patents with underlying diseases, primarily haematological malignancies (57/72 [80%]). Blood cultures were positive in 70% of patients suffering from disseminated mycosis. Neutropenia, fever and cerebral symptoms were independently related to the development of disseminated infection whereas recovery from aplasia was associated with a reduced risk. The overall mortality was 46.9% but mortality rate was 87.5% in patients with disseminated disease. Survival was independently associated with surgical excision and recovery from aplasia. Antifungal treatments were not related to a reduced risk of death. Infections caused by S. prolificans are life threatening in susceptible patients, and can be considered a truly emerging disease. Infections are difficult to treat since it is a multi-resistant species. Multicenter studies are essential with the aim of developing and disseminating appropriate techniques and protocols to treat this mycosis.

Differences in pathogenicity and clinical syndromes due to Aspergillus fumigatus and Aspergillus flavus

Abstract: Most of the information available about Aspergillus infections has originated from the study of A. fumigatus, the most frequent species in the genus. This review aims to compare the pathogenicity and clinical aspects of Aspergillosis caused by A. fumigatus an A. flavus. Experimental data suggests that A. flavus is more virulent than A. fumigatus. However, these were mostly models of disseminated Aspergillus infection which do not properly mimic the physiopathology of invasive aspergillosis, a condition that is usually acquired by inhalation. In addition, no conclusive virulence factor has been identified for Aspergillus species. A. flavus is a common cause of fungal sinusitis and cutaneous infections. Chronic conditions such as chronic cavitary pulmonary aspergillosis and sinuses fungal balls have rarely been associated with A. flavus. The bigger size of A. flavus spores, in comparison to those of A. fumigatus spores, may favour their deposit in the upper respiratory tract. Differences between these species justify the need for a better understanding of A. flavus infections.

Metabolomics of Aspergillus fumigatus.

Abstract: Aspergillus fumigatus is the most important species in Aspergillus causing infective lung diseases. This species has been reported to produce a large number of extrolites, including secondary metab...

Multiple-azole-resistant Aspergillus fumigatus osteomyelitis in a patient with chronic granulomatous disease successfully treated with long-term oral posaconazole and surgery

Abstract: We describe a patient with chronic granulomatous disease and proven Aspergillus fumigatus osteomyelitis of the midfoot, while receiving itraconazole-prophylaxis. The isolate proved resistant to itraconazole as well as voriconazole, and showed reduced susceptibility to posaconazole. Although molecular analysis demonstrated the presence of a 53 base pair tandem repeat in the promoter region for cyp51A, i.e., the gene coding for the target enzyme of the azole antifungals, there were no mutations in the cyp51A gene. Since transformation of the promoter region into wild-type strains did not result in an azole resistant phenotype, a yet unknown mutation was suspected. The patient was treated with extensive surgery and two weeks of caspofungin therapy, followed by one year of posaconazole therapy. He made a complete recovery and did not experience any side effects. Long-term posaconazole proved to be a safe and effective treatment for multi-azole resistant A. fumigatus osteomyelitis in this immunocompromised patient.

Role of (1→3)-β-D-glucan in the diagnosis of invasive aspergillosis

Abstract: Measurement of serum (1-->3)-beta-D-Glucan (BG) is an aid in the diagnosis of fungemia and deep-seated mycoses, including invasive aspergillosis (IA). BG is present in the cell wall of most pathogenic fungi (including Pneumocystis jiroveci) in significant amounts with some notable exceptions such as Cryptococcus neoformans and Zygomycetes. Commercially available assays can detect serum BG concentrations as low as 1 pg/mL. Published validation studies have included patients with IA and other invasive fungal diseases (IFD). BG detection appears to be more sensitive than galactomannan detection in patients with IA, but BG's intrinsic lack of mycological specificity requires the integration of clinical, radiological, and microbiological data for proper interpretation. BG assay test characteristics can be used, for example, to exclude IA in some clinical scenarios, to increase the certainty of IA in the presence of an isolated positive galactomannan result or when testing follows initiation of antifungal treatment. BG may be falsely elevated in the serum in the absence of IFD in patients undergoing hemodialysis with cellulose membranes, in patients treated with immunoglobulin, albumin, or other blood products filtered through cellulose filters containing BG, and in patients with serosal exposure to glucan-containing gauze or to certain intravenous antimicrobials. These potential sources of false positivity should be considered when interpreting BG results. BG may be useful as a sensitive screening tool for surveillance of IA and other IFD in populations at risk. Stratified IFD screening and diagnostic strategies using both galactomannan and BG should be explored. Factors affecting the production and clearance of BG during IA and other IFD need additional study to further refine its diagnostic utility.

The characteristics of Aspergillus fumigatus mycetoma development: is this a biofilm?

Abstract: Aspergillus fumigatus is an increasingly prevalent opportunistic fungal pathogen of various immuno-compromised individuals. It has the ability to filament within the lungs forming dense intertwined mycelial balls. These morphological characteristics resemble those of microbial biofilms, which are matrix enclosed microbial populations, adherent to each other and/or to surfaces or interfaces. The purpose of this paper is to review some recent experiments that indicate the potential biofilm forming capacity of A. fumigatus in vitro. Initially it was established that conidial seeding density is important for stable biofilm development. In the optimized model conidial germination and filamentous growth characteristics were not observed until 8 h, after which a multi-cellular population expanded exponentially forming a thick structure (approx. 250 microm). Calcofluor white staining of this revealed the presence of extracellular polymeric matrix material, which increased as the biofilm matured. Subsequent antifungal sensitivity testing of this structure showed that azoles, polyenes and echinocandins were ineffective in reducing the cellular viability at therapeutically attainable concentrations. Microarray and real-time PCR analysis demonstrated the up-regulation of AfuMDR4 during multicellular growth and development, which may account the recalcitrance observed. Overall, A. fumigatus appears to possess the classical elements of biofilm growth, namely multicellularity, matrix production and sessile resistance. This controversial approach to understanding the biology of A. fumigatus infection may provide crucial information on how to treat this pathogenic fungus more effectively.

Allergic fungal sinusitis: pathophysiology, diagnosis and management.

Abstract: Allergic fungal sinusitis (AFS) is a noninvasive form of fungal rhinosinusitis with an incidence of between 6 and 9% of all rhinosinusitis requiring surgery. Regional variation in incidence has been reported, with the southern and southwestern US particularly endemic. Patients with AFS commonly present with chronic rhinosinusitis with nasal polyps, inhalant atopy, elevated total serum immunoglobulin E (IgE), and sinus-obstructing inspissates of a characteristic extramucosal 'peanut buttery' visco-elastic eosinophil-rich material called 'allergic mucin' that contains sparse numbers of fungal hyphae. Sinus CT is always abnormal, showing findings of chronic rhinosinusitis that often include central areas of increased contrast ('hyperattenuation') within abnormal paranasal sinuses that represent the presence of fungal-containing allergic mucin. AFS has been found to be analogous in several ways to allergic bronchopulmonary aspergillosis (ABPA). Both are chronic inflammatory respiratory tract disorders that are driven by hypersensitivity responses to the presence of small numbers of extramucosal fungi found growing within airway-impacting allergic mucin. AFS allergic mucin typically cultures positive for either dematiaceous fungi such as Bipolaris spicifera or Curvularia lunata, or Aspergillus species such as A. fumigatus, A. flavus or A. niger. As with ABPA, patients have type I immediate hypersensitivity to the etiologic mold in AFS. Further, both AFS and ABPA have been found to have association with specific class II major histocompatibility alleles. Proper diagnosis of AFS and differentiation from the other forms of both noninvasive and invasive fungal rhinosinusitis requires strict adherence to published diagnostic criteria. Medical treatment of AFS has been modeled to an extent after treatment approaches for ABPA that includes the use of postoperative oral corticosteroids and aggressive antiallergic inflammation therapy. The use of follow-up measurements of total serum IgE during treatment of both AFS and ABPA patients can help to monitor disease activity. Future AFS research will lead to further insights into pathogenesis, improved treatments, and ultimately decreases in surgical recurrence rates for this highly recurrent hypertrophic rhinosinusitis disorder.

What do we know about the role of gliotoxin in the pathobiology of Aspergillus fumigatus

Abstract: Gliotoxin is a member of the epipolythiodioxopiperazine class of toxins and is both the major and the most potent toxin produced by Aspergillus fumigatus. Since the discovery of the putative gliotoxin biosynthetic 12-gene cluster in the genome of A. fumigatus, five different laboratories have attempted to determine the role of this toxin in the virulence of A. fumigatus. The genes in the cluster that have been disrupted to study the pathobiological importance of gliotoxin include gliZ that encodes a transcription factor and gliP that encodes a nonribosomal peptide synthase. Two of the five laboratories have reported gliotoxin to be an important virulence determinant of A. fumigatus, while the other three laboratories have shown it to be unimportant. Comparisons of the data generated among the five laboratories revealed that the immunosuppressive regimen used for mice was the key factor that contributed to the observed disparity. Regardless of either the mouse strains used or the route of infection, immunosuppression with a combination of cyclophosphamide and corticosteroids (neutropenic mice) showed gliotoxin to be unimportant. The mice immunosuppressed with corticosteroids alone, however, revealed that gliotoxin is an important virulence determinant of A. fumigatus. These studies indicate that the neutropenic mice model is inadequate to reveal the pathobiological importance of fungal secondary metabolites in invasive pulmonary aspergillosis.

Ecology of Pseudallescheria and Scedosporium species in human-dominated and natural environments and their distribution in clinical samples.

Abstract: This study aims to determine the occurrence of Pseudallescheria and Scedosporium species in natural and human-dominated environments. Habitats (136 sampling sites) in a transect with increasing human impact were investigated (natural areas, agricultural soils, urban playgrounds, industrial areas). Physico-chemical parameters were measured to characterize the different areas included in this investigation. Fungal identification was performed by morphology and sequence data analysis. Comparative description of virulence was largely based on the database of the ECMM/ISHAM Working Group on Pseudallescheria/Scedosporium Infections. Pseudallescheria and Scedosporium species were most abundant in industrial areas, followed by urban playgrounds and agricultural areas. None of the species were isolated from natural habitats. The abundance of Pseudallescheria and Scedosporium species could be correlated with increasing nitrogen concentrations (P<0.01) and decreasing pH (P<0.05) within a pH range of 6.1-7.5. In general, frequency of the different Pseudallescheria and Scedosporium species in the environment is strongly enhanced by human activities, and largely differs from species distribution in clinical settings, suggesting that these species have different degrees of virulence. Pseudallescheria boydii is relatively frequently found as agent of human disease, while Scedosporium dehoogii is found almost exclusively in the environment. Scedosporium apiospermum is responsible for the majority of infections and is found at comparable frequency in the environment; S. aurantiacum and P. minutispora showed similar spectra, but at much lower frequencies.

In vitro antifungal activities of luliconazole, a new topical imidazole.

Abstract: Luliconazole is a topical antifungal drug newly developed in Japan. The present study compares the in vitro antifungal activity of luliconazole against clinically important dermatomycotic fungi with that of other representative antifungal drugs. The reference drugs chosen were five classes of nine topical agents, i.e., allylamine (terbinafine), thiocarbamate (liranaftate), benzylamine (butenafine), morpholine (amorolfine), and azole (ketoconazole, clotrimazole, neticonazole, miconazole and bifonazole). The minimum inhibitory concentrations (MIC) of luliconazole and the reference drugs against Trichophyton spp. (T. rubrum, T. mentagrophytes and T. tonsurans) and Candida albicans were measured by the standardized broth microdilution method. Luliconazole demonstrated greater potency against Trichophyton spp. (MIC range:

What is a species in Aspergillus

Abstract: The various species concepts with emphasis on those which can be applied to Aspergillus and its teleomorphs are discussed. Any proposed new species should show evidence for evolutionary divergence from other taxa, particularly unique DNA characters at multiple loci and the polyphasic approach was suggested as the 'gold standard' for species delimitation using a combination of multilocus sequence data, morphological, physiological characteristics and ecological data. For species descriptions it is recommended to examine several gene sequences (e.g., ITS, calmodulin, beta-tubulin, actin) and submit them to recognized sequence databases. Dual naming of Aspergillus taxa with teleomorphs has been recommended where necessary. To avoid confusion, the 'List of Names in Current Use' is recommended as a reference for Aspergillus nomenclature. For clinical researchers who depend on one of the names, it was suggested to use the name 'complex' if identification is solely based on morphology, which cannot distinguish between closely related species. A protocol for naming new Aspergillus taxa is proposed, which include the deposition of type cultures in at least two recognized culture collections. If type cultures are not available the taxon can be declared invalid.

Th17 cells in the setting of Aspergillus infection and pathology

Abstract: Innate and adaptive immune responses act to generate the most effective form of immunity for protection against Aspergillus fumigatus. The decision of how to respond is still primarily determined by interactions between fungi and cells of the innate immune system, but the actions of T cells will feed back into this dynamic equilibrium to regulate the balance between pro-inflammatory and anti-inflammatory signals. The enzyme indoleamine 2,3-dioxygenase, and tryptophan metabolites, acting as a bridge between dendritic cells and regulatory T cells, pivotally contribute to such a homeostatic condition by taming inflammatory responses. IL-23 and the newly described Th17 pathway, by means of negative regulation of tryptophan catabolism, play an inflammatory role previously attributed to uncontrolled Th1 response. Our data support a model in which IL-23/IL-17A/Th17-driven inflammation promotes infection and impairs antifungal immune resistance. Thus, modulation of the inflammatory response represents a potential strategy to stimulate protective immune responses to Aspergillus.

Invasive aspergillosis in chronic granulomatous disease

Abstract: Chronic granulomatous disease (CGD) is a rare inherited disorder of the NADPH oxidase complex in which phagocytes are defective in generating superoxide anion. NADPH oxidase activation leads to release of sequestered neutrophil granular proteases, which are likely the principal antimicrobial effectors. CGD is characterized by recurrent life-threatening bacterial and fungal infections and by inflammatory complications such as wound dehiscence, obstructive granulomata of the genitourinary tract, and inflammatory bowel disease. Despite routine use of interferon-gamma prophylaxis, fungal infections have remained a persistent problem with an incidence of 0.1 fungal infections per patient year. Invasive pulmonary aspergillosis in CGD manifests with neutrophils and lymphohistiocytic inflammation; hyphal angioinvasion is not observed, indicating that NADPH oxidase-independent pathways appear to be adequate to protect against vascular invasive disease. Chronic prophylaxis with a mould-active agent is standard of care in CGD. “Mulch pneumonitis” is characterized by rapid onset life- pulmonary inflammation following mould exposure, and treated with systemic antifungals and prolonged corticosteroids. Haematopoietic stem cell transplantation is curative in CGD, but transplant-related mortality from GVHD and infectious complications is substantial, particularly in cases of donor/recipient HLA-antigen disparity. Since CGD is a disorder of myeloid stem cells, gene therapy is an attractive option, but maintaining stable population of myeloid gene-corrected cells has been a persistent challenge. The strategy of non-myeloablative conditioning followed by gene therapy facilitates the expansion of gene-corrected cells. Studies of experimental aspergillosis in CGD point to defective tryptophan metabolism as a contributing factor to impaired host defence and increased lung inflammation in CGD.

Cryptococcosis in domestic animals in Western Australia: a retrospective study from 1995–2006

Abstract: A retrospective study of cryptococcosis in domestic animals residing in Western Australia was conducted over an 11-year-period (from 1995 to 2006) by searching the data base of Murdoch University Veterinary Teaching hospital and the largest private clinical pathology laboratory in Perth. Cryptococcosis was identified in 155 animals: 72 cats, 57 dogs, 20 horses, three alpacas, two ferrets and a sheep. There was no seasonal trend apparent from the dates of diagnosis. Taking into account the commonness of accessions to Murdoch University, cats were five to six times more likely to develop this disease than dogs, and three times more likely than horses, while horses were almost twice as likely as dogs to become infected. Amongst the feline cohort, Ragdoll and Birman breeds were over-represented, while in dogs several pedigree breeds were similarly overrepresented. Dogs and horses tended to develop disease at an early age (one to five years), while cats were presented over a much wider range of ages. In cats and dogs the upper respiratory tract was the most common primary site of infection, while horses and alpacas tended to have lower respiratory involvement. The most striking finding of the study was the high frequency with which C. gattii was identified, with infections attributable to this species comprising 5/9 cats, 11/22 dogs, 9/9 horses and 1/1 alpaca, where appropriate testing was conducted. Preliminary molecular genotyping suggested that most of the C. gattii infections in domestic animals (9/9 cases) were of the VGII genotype. This contrasts the situation on the eastern seaboard of Australia, where disease attributable to C. gattii is less common and mainly due to the VGI genotype. C. gattii therefore appears to be an important cause of cryptococcosis in Western Australia.

Detection of 2-Pentylfuran in the breath of patients with Aspergillus fumigatus

Abstract: Aspergillus fumigatus produces 2-Pentylfuran (2-PF) when cultured on blood agar, nutrient agar and other media. As 2-PF is not known to be produced by mammalian metabolism we hypothesized that it is detectable in breath of patients colonized or infected with A. fumigatus. Breath was tested for 2-PF from normal subjects, those undergoing chemotherapy, and adults at risk of colonization or infection with A. fumigatus because of bronchiectasis, cystic fibrosis, or immune suppression. Breath samples were collected in five L tedlar bags and analyzed by Gas Chromatography/Mass Spectroscopy (GC/MS) in MS-MS mode. 2-PF was not detected in breath 14 healthy controls, in one of 10 neutropenic subjects and 16 of 32 patients with lung disease. The sensitivity and specificity of the 2-PF breath tests when compared with recurrent isolation of aspergillus from sputum or from bronchoalveolar lavage over two months was 77% and 78% respectively. As 2-PF is not normally found in human breath its presence may reflect the act...

TLR-2, TLR-4 and dectin-1 expression in human monocytes and neutrophils stimulated by Paracoccidioides brasiliensis.

Abstract: Paracoccidioidomycosis (PCM) is an endemic mycosis in Latin America caused by the dimorphic fungus Paracoccidioides brasiliensis. The pattern of the immune responses to P. brasiliensis determines the disease progression and clinical outcome. Innate immune response is mediated by phagocytic cells, such as macrophage and neutrophils, which ingest and kill invading pathogens and then trigger the adaptive immune system through the secretion of cytokines and chemokines. The C-type like lectin receptors (CLR) and Toll-like receptors (TLRs) are the two main pattern recognition receptors in phagocytic cells that recognize fungal components. Therefore, the purpose of the present study was to evaluate the expression of TLR-1, TLR-2, TLR-4 and dectin-1 (CLR) in monocytes and neutrophils from healthy individuals after stimulation with Pb18 (high virulence) and Pb265 (low virulence) yeasts of P. brasiliensis. As positive controls we used specific ligands to TLR-4 (LPS), TLR-2 and dectin-1 (zymosan). Our results demonstrated a decreased of TLRs and dectin-1 expression mainly on monocytes as opposes on neutrophils, as soon as 30 minutes after yeast cells stimulation. This decrease was similar to the one caused by zymosan stimulation and indicates that up binding the complexes are rapidly internalized. There was a tendency towards an increased TLR2 and dectin-1 mRNA expression in response to fungal cells, mainly to Pb265. P. brasiliensis yeast cells induced the production of pro-inflammatory and anti-inflammatory cytokines, but the low ratio between TNF-alpha and IL-10 in response to zymosan and Pb265 indicates a balanced production of IL-10 and TNF-alpha, while Pb18 predominantly induced TNF-alpha secretion. Fungal cells also induced an elevated production of PGE(2) by monocytes and neutrophils showing their potential to provoke an intense inflammatory response. Altogether our results suggest the participation of TLR2, TLR-4 and dectin-1 in P. brasiliensis recognition, internalization and consequent activation of the immune response against the fungus. Moreover, the preferential recognition of zymosan and Pb265 by TLR-2 and dectin-1 would result in the production of adequate concentration of IL-10, which would be able to counterbalance the excessive inflammatory response mediated by TNF-alpha and PGE2. With these attributes the low virulence strain of P. brasiliensis would induce a controlled immune response beneficial to the host.

Invasive aspergillosis in glucocorticoid-treated patients

Abstract: Glucocorticoids have potent, pleiotropic effects on the immune system that can predispose patients to developing life-threatening invasive aspergillosis (IA). While the exact prevalence and attributable mortality of IA in glucocorticoid-treated patients is difficult to estimate, Aspergillus species are significant pathogens in patients that require prolonged high-dose glucocorticoid therapy including multiple myeloma, collagen vascular diseases, or recipients of solid organ/hematopoietic transplantation. Experimental animal models and autopsy series have revealed important differences in the pathology of aspergillosis between glucocorticoid-treated and neutropenic patients. Although neutropenic hosts develop infection characterized by extensive angioinvasion, hemorrhagic thrombosis and necrosis with a high fungal burden, glucocorticoid-immunosuppressed hosts present with infection dominated by extensive necrosis, less angioinvasion, and a lower fungal burden suggestive of an inflammation-driven pathology. These pathobiological differences may have important implications for the diagnosis and treatment approaches of IA in glucocorticoid-treated patients.

Fungal strategies for overcoming host innate immune response.

Abstract: A successful pathogen is one that is able to effectively survive and evade detection by the host innate immune defense. Fungal pathogens have adopted strategies which evade host defense and eventually cause disease in at-risk patients. Shielding of stimulatory surface recognition molecules, shedding of decoy components, induction of anti-inflammatory signals, complement evasion and resilient survival capacity are successful evasion mechanisms employed by fungal pathogens. Understanding these complex pathways of immune evasion can potentially contribute to development of novel therapeutic strategies against fungal infections.

Sexual structures in Aspergillus: morphology, importance and genomics.

Abstract: The genus Aspergillus comprises a few hundred species sharing a common asexual spore forming structure, the aspergillum. Approximately one-third of these species also produce a sexual stage, all bu...

Immune dysfunction in Atlantic bottlenose dolphins (Tursiops truncatus) with lobomycosis

Abstract: Lobomycosis (Lacaziosis) occurs only in humans and dolphins under natural conditions. We evaluated the immune status of eight dolphins with lobomycosis and 40 healthy dolphins from the Indian River Lagoon (IRL), Florida. Lobomycosis cases had multiple abnormalities in their immunologic parameters when compared to healthy dolphins. The absolute number of circulating lymphocytes and serum albumin concentration were reduced (P<0.05) while the segmented neutrophils, alpha 1, total beta, total gamma and total globulins were increased (P<0.05). Although innate immunity was relatively intact and phagocytosis and natural killer cell activity were not affected, the plasma lysozyme concentrations were elevated in dolphins with lobomycosis (P<0.05). Adaptive immunity was depressed with statistically significant decreases found in the absolute numbers of CD4(+) helper T cells and CD19(+) and CD21(+) B cells. The ratios of CD2(+) T cells to CD4(+) cells and CD2(+) to CD21(+) cells were increased (P=0.05 and P<0.05, respectively) and the numbers of lymphocytes expressing MHC class II molecules was decreased in dolphins with lobomycosis (P<0.05). Lymphocyte proliferation was reduced in response to stimulation with lipopolysaccharide and concanavalin A (P<0.05). Antibody titers to Erysipelas rhusiopathiae, a common marine micro-organism, were decreased (P<0.05). In summary, dolphins with lobomycosis exhibit significant impairment in adaptive immunity.

Biofilm formation on intrauterine devices in patients with recurrent vulvovaginal candidiasis

Abstract: A biofi lm is a complex community of surface-associated cells enclosed in a polymer matrix. They attach to solid surfaces and their formation can be affected by growth conditions and co-infection with other pathogens. The presence of biofi lm may protect the microorganisms from host defenses, as well as signifi cantly reduce their susceptibility to antifungal agents. Pathogenic microbes can form biofi lms on the inert surfaces of implanted devices such as catheters, prosthetic cardiac valves and intrauterine devices (IUDs). The present study was carried out to analyze the presence of biofi lm on the surface of intrauterine devices in patients with recurrent vulvovaginal candidiasis, and to determine the susceptibility profi le of the isolated yeasts to amphotericin B and fl uconazole. Candida albicans was recovered from the IUDs and it was found to be susceptible to the antifungal agents when tested under planktonic growing conditions. These fi ndings indicate the presence of the biofi lm on the surface of the IUD as an important risk factor for recurrent vulvovaginal candidiasis.

Comparison of techniques to examine the diversity of fungi in adult patients with cystic fibrosis.

Abstract: This study compares conventional and molecular techniques for the detection of fungi in 77 adult cystic fibrosis (CF) patients. Three different methods were investigated, i.e., (1) conventional microbiological culture (including yeasts and filamentous fungi), (2) mycological culture with CF-derived fungal specific culture media, and (3) Non-culture and direct DNA extraction from patient sputa. Fungi isolated from environmental air samples of the CF unit were compared to fungi in sputa from CF patients. Fungi (n = 107) were detected in 14/77(18%) of patients by method 1, in 60/77 (78%) of patients by method 2 and with method 3, in 77/77(100%) of the patients. The majority of yeasts isolated were Candida albicans and C. dubliniensis. Exophiala (Wangiella) dermatitidis, Scedosporium apiospermum, Penicillium spp., Aspergillus fumigatus, and Aspergillus versicolor were also identified by sequence analysis of the rDNA short internal transcribed spacer (ITS2) region. Conventional laboratory analysis failed to de...

Cerebral aspergillosis: tissue penetration is the key

Abstract: Cerebral aspergillosis is increasingly recognized in severely immunocompromised patients and, until recently, this type of fungal infection was associated with a mortality approaching 100%. The central nervous system is a protected environment and penetration of drugs across the blood-brain barrier is mainly limited by their molecular size and physicochemical properties, as well as drug interaction with transporter systems (e.g., P-glycoprotein) at the blood-brain barrier. Most antifungal agents are large molecules (� 700 Da), which makes sufficient penetration into the central nervous system unlikely. In fact, the available data indicate low levels of most antifungal agents in cerebrospinal fluid and brain tissue, except for fluconazole and voriconazole. Concentrations of voriconazole exceeding inhibitory concentrations for Aspergillus species were found repeatedly in cerebrospinal fluid and brain tissue, including brain abscess material. A recent retrospective study confirmed that voriconazole treatment resulted in improved response and survival rates in patients with cerebral aspergillosis. Data from animal models, which explored escalated doses or combinations of antifungal agents in experimental neuroaspergillosis, suggest that selected combination or doseescalated therapies might further improve the still unsatisfactory prognosis in this particular type of Aspergillus infection.

In vitro activities of isavuconazole against opportunistic filamentous and dimorphic fungi

Abstract: The in vitro activity of isavuconazole was compared to those of amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole, and ravuconazole against 300 clinical isolates of Pseudallescheria boydii, Paecilomyces lilacinus, Fusarium spp., Bipolaris spicifera, Curvularia lunata, Alternaria alternata, Exophiala spp., Rhizopus arrhizus, Mucor circillenoides, Absidia corymbifera, Blastomyces dermatitidis, Histoplasma capsulatum and Coccidioides posadasii. MICs were determined by a broth macrodilution method based on the CLSI M38-A procedure. The triazoles were relatively uniform in that they showed strong in vitro inhibitory activity against most of the tested fungi. In vitro activity was variable with strains of P. lilacinus while with Fusarium spp., the triazoles were found to have limited in vitro activity and amphotericin B was moderately active. The results suggest that isavuconazole is a broad-spectrum antifungal agent, effective against a wide range of moulds in vitro.

Galactofuranose containing molecules in Aspergillus fumigatus

Abstract: Galactofuranose is a major carbohydrate in Aspergillus fumigatus. It became famous in medical mycology as being part of the galactomannan which was shown 30 years ago to be the major antigen circulating in the body fluid of patients suffering from invasive aspergillosis. Four different molecules contain galactofuranose in A. fumigatus: (i) the galactomannan present in the alkali soluble and insoluble fraction of the cell wall (ii) N- and O glycan moieties of secreted glycoproteins (iii) a GPI- anchored lipophosphogalactomannan and (iv) several sphingolipids also anchored to the membrane by an inositol phosphoceramide.

Genetic diversity and species delimitation in the opportunistic genus Fonsecaea

Abstract: Genetic diversity and species delimitation were investigated among 39 isolates recovered from clinical and environmental sources in Central and South America, Africa, East Asia and Europe. All had been morphologically identified as Fonsecaea spp. Molecular analyses were based on sequences of the ribosomal internal transcribed spacers (ITS), -tubulin (TUB1) and actin (ACT1) regions. A phylogenetic approach using haplotype networks was used to evaluate species delimitation and genetic diversity. The presence and the modes of reproductive isolation were tested by measuring the index of differentiation (ID) and the index of association (IA). Based on the sequence data, the 39 Fonsecaea strains were classified into three major entities: (i) a group representing Fonsecaea pedrosoi, (ii) a second composed of F. monophora, and (iii) a third group including mostly strains from South America. The two major, clinically relevant Fonsecaea species, F. monophora and F. pedrosoi, also differed in the pathological symptoms found in patients. Moreover, F. pedrosoi is mostly recovered in clinical settings, whereas F. monophora is commonly isolated from the environment. One environmental strain with Fonsecaea-like appearance was shown to belong to a different species, only distantly related to the core-group of Fonsecaea.