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J. Fungi, Volume 5, Issue 2 (June 2019)

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Cover Story (view full-size image) Low virulence Candida species can provide protection against lethal polymicrobial sepsis caused by [...] Read more.
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Open AccessArticle
Aspergillus flavus NRRL 35739, a Poor Biocontrol Agent, May Have Increased Relative Expression of Stress Response Genes
J. Fungi 2019, 5(2), 53; https://doi.org/10.3390/jof5020053
Received: 4 May 2019 / Revised: 16 June 2019 / Accepted: 18 June 2019 / Published: 20 June 2019
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Abstract
Biocontrol of the mycotoxin aflatoxin utilizes non-aflatoxigenic strains of Aspergillus flavus, which have variable success rates as biocontrol agents. One non-aflatoxigenic strain, NRRL 35739, is a notably poor biocontrol agent. Its growth in artificial cultures and on peanut kernels was found to [...] Read more.
Biocontrol of the mycotoxin aflatoxin utilizes non-aflatoxigenic strains of Aspergillus flavus, which have variable success rates as biocontrol agents. One non-aflatoxigenic strain, NRRL 35739, is a notably poor biocontrol agent. Its growth in artificial cultures and on peanut kernels was found to be slower than that of two aflatoxigenic strains, and NRRL 35739 exhibited less sporulation when grown on peanuts. The non-aflatoxigenic strain did not greatly prevent aflatoxin accumulation. Comparison of the transcriptomes of aflatoxigenic and non-aflatoxigenic A. flavus strains AF36, AF70, NRRL 3357, NRRL 35739, and WRRL 1519 indicated that strain NRRL 35739 had increased relative expression of six heat shock and stress response proteins, with the genes having relative read counts in NRRL 35739 that were 25 to 410 times more than in the other four strains. These preliminary findings tracked with current thought that aflatoxin biocontrol efficacy is related to the ability of a non-aflatoxigenic strain to out-compete aflatoxigenic ones. The slower growth of NRRL 35739 might be due to lower stress tolerance or overexpression of stress response(s). Further study of NRRL 35739 is needed to refine our understanding of the genetic basis of competitiveness among A. flavus strains. Full article
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Open AccessReview
Mass Spectrometry-Based Proteomics of Fungal Pathogenesis, Host–Fungal Interactions, and Antifungal Development
J. Fungi 2019, 5(2), 52; https://doi.org/10.3390/jof5020052
Received: 1 June 2019 / Revised: 12 June 2019 / Accepted: 14 June 2019 / Published: 17 June 2019
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Abstract
The prevalence of fungal diseases is increasing on a global scale, ranging from acute to systemic infections caused by commensal or pathogenic microorganisms, often associated with the immune status of the host. Morbidity and mortality rates remain high and our ability to treat [...] Read more.
The prevalence of fungal diseases is increasing on a global scale, ranging from acute to systemic infections caused by commensal or pathogenic microorganisms, often associated with the immune status of the host. Morbidity and mortality rates remain high and our ability to treat fungal infections is challenged by a limited arsenal of antifungal agents and the emergence of drug resistant pathogens. There is a high demand for new approaches to elucidate the fungal mechanisms of pathogenesis and the interplay between host and pathogen to discover novel treatment options. Moreover, the need for improved drug efficacy and reduced host toxicity requires the identification and characterization of antifungal biological targets and molecular mechanisms of action. Mass spectrometry (MS)-based proteomics is a rapidly advancing field capable of addressing these priorities by providing comprehensive information on the dynamics of cellular processes, modifications, and interactions. In this Review, we focus on applications of MS-based proteomics in a diverse array of fungal pathogens and host systems to define and distinguish the molecular details of fungal pathogenesis and host–fungal interactions. We also explore the emerging role of MS-based proteomics in the discovery and development of novel antifungal therapies and provide insight into the future of MS-based proteomics in fungal biology. Full article
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Open AccessCommentary
The Fight against HIV-Associated Disseminated Histoplasmosis in the Americas: Unfolding the Different Stories of Four Centers
J. Fungi 2019, 5(2), 51; https://doi.org/10.3390/jof5020051
Received: 14 May 2019 / Revised: 13 June 2019 / Accepted: 14 June 2019 / Published: 17 June 2019
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Abstract
Disseminated histoplasmosis is a major opportunistic infection of HIV-infected patients, killing thousands in Latin America each year. Yet, it remains a neglected disease that is often confused with tuberculosis, for lack of simple, affordable, and rapid diagnostic tools. There is great heterogeneity in [...] Read more.
Disseminated histoplasmosis is a major opportunistic infection of HIV-infected patients, killing thousands in Latin America each year. Yet, it remains a neglected disease that is often confused with tuberculosis, for lack of simple, affordable, and rapid diagnostic tools. There is great heterogeneity in the level of histoplasmosis awareness. The purpose of this report was to describe how the historical “awakening” to the threat of histoplasmosis came to be in four different centers that have actively described this disease: In Brazil, the Sao José hospital in Fortaleza; in Colombia, the Corporación para Investigaciones Biológicas in Medellin; in French Guiana, Cayenne Hospital; and in Guatemala, the Association de Salud Integral in Guatemala city. In Brazil and French Guiana, the search for leishmaniasis on the buffy coat or skin smears, respectively, led to the rapid realization that HIV patients were suffering from disseminated histoplasmosis. With time and progress in fungal culture, the magnitude of this problem turned it into a local priority. In Colombia and Guatemala, the story is different because for these mycology centers, it was no surprise to find histoplasmosis in HIV patients. In addition, collaborations with the CDC to evaluate antigen-detection tests resulted in researchers and clinicians developing the capacity to rapidly screen most patients and to demonstrate the very high burden of disease in these countries. While the lack of awareness is still a major problem, it is instructive to review the ways through which different centers became histoplasmosis-aware. Nevertheless, as new rapid diagnostic tools are becoming available, their implementation throughout Latin America should rapidly raise the level of awareness in order to reduce the burden of histoplasmosis deaths. Full article
(This article belongs to the Special Issue Histoplasma and Histoplasmosis)
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Open AccessReview
A New Front in Microbial Warfare—Delivery of Antifungal Effectors by the Type VI Secretion System
J. Fungi 2019, 5(2), 50; https://doi.org/10.3390/jof5020050
Received: 17 May 2019 / Revised: 11 June 2019 / Accepted: 13 June 2019 / Published: 14 June 2019
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Abstract
Microbes typically exist in mixed communities and display complex synergistic and antagonistic interactions. The Type VI secretion system (T6SS) is widespread in Gram-negative bacteria and represents a contractile nano-machine that can fire effector proteins directly into neighbouring cells. The primary role assigned to [...] Read more.
Microbes typically exist in mixed communities and display complex synergistic and antagonistic interactions. The Type VI secretion system (T6SS) is widespread in Gram-negative bacteria and represents a contractile nano-machine that can fire effector proteins directly into neighbouring cells. The primary role assigned to the T6SS is to function as a potent weapon during inter-bacterial competition, delivering antibacterial effectors into rival bacterial cells. However, it has recently emerged that the T6SS can also be used as a powerful weapon against fungal competitors, and the first fungal-specific T6SS effector proteins, Tfe1 and Tfe2, have been identified. These effectors act via distinct mechanisms against a variety of fungal species to cause cell death. Tfe1 intoxication triggers plasma membrane depolarisation, whilst Tfe2 disrupts nutrient uptake and induces autophagy. Based on the frequent coexistence of bacteria and fungi in microbial communities, we propose that T6SS-dependent antifungal activity is likely to be widespread and elicited by a suite of antifungal effectors. Supporting this hypothesis, homologues of Tfe1 and Tfe2 are found in other bacterial species, and a number of T6SS-elaborating species have been demonstrated to interact with fungi. Thus, we envisage that antifungal T6SS will shape many polymicrobial communities, including the human microbiota and disease-causing infections. Full article
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Open AccessArticle
Integrated Analysis of Clinical and Microbiome Risk Factors Associated with the Development of Oral Candidiasis during Cancer Chemotherapy
J. Fungi 2019, 5(2), 49; https://doi.org/10.3390/jof5020049
Received: 27 May 2019 / Revised: 11 June 2019 / Accepted: 12 June 2019 / Published: 13 June 2019
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Abstract
Oral candidiasis is a common side effect of cancer chemotherapy. To better understand predisposing factors, we followed forty-five subjects who received 5-fluorouracil- or doxorubicin-based treatment, during one chemotherapy cycle. Subjects were evaluated at baseline, prior to the first infusion, and at three additional [...] Read more.
Oral candidiasis is a common side effect of cancer chemotherapy. To better understand predisposing factors, we followed forty-five subjects who received 5-fluorouracil- or doxorubicin-based treatment, during one chemotherapy cycle. Subjects were evaluated at baseline, prior to the first infusion, and at three additional visits within a two-week window. We assessed the demographic, medical and oral health parameters, neutrophil surveillance, and characterized the salivary bacteriome and mycobiome communities through amplicon high throughput sequencing. Twenty percent of all subjects developed oral candidiasis. Using multivariate statistics, we identified smoking, amount of dental plaque, low bacteriome and mycobiome alpha-diversity, and the proportions of specific bacterial and fungal taxa as baseline predictors of oral candidiasis development during the treatment cycle. All subjects who developed oral candidiasis had baseline microbiome communities dominated by Candida and enriched in aciduric bacteria. Longitudinally, oral candidiasis was associated with a decrease in salivary flow prior to lesion development, and occurred simultaneously or before oral mucositis. Candidiasis was also longitudinally associated with a decrease in peripheral neutrophils but increased the neutrophil killing capacity of Candida albicans. Oral candidiasis was not found to be associated with mycobiome structure shifts during the cycle but was the result of an increase in Candida load, with C. albicans and Candida dubliniensis being the most abundant species comprising the salivary mycobiome of the affected subjects. In conclusion, we identified a set of clinical and microbiome baseline factors associated with susceptibility to oral candidiasis, which might be useful tools in identifying at risk individuals, prior to chemotherapy. Full article
(This article belongs to the Special Issue Oral Mycobiome)
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Open AccessReview
Interactions between Aspergillus fumigatus and Pulmonary Bacteria: Current State of the Field, New Data, and Future Perspective
J. Fungi 2019, 5(2), 48; https://doi.org/10.3390/jof5020048
Received: 18 April 2019 / Revised: 10 June 2019 / Accepted: 10 June 2019 / Published: 12 June 2019
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Abstract
Aspergillus fumigatus and Pseudomonas aeruginosa are central fungal and bacterial members of the pulmonary microbiota. The interactions between A. fumigatus and P. aeruginosa have only just begun to be explored. A balance between inhibitory and stimulatory effects on fungal growth was observed in [...] Read more.
Aspergillus fumigatus and Pseudomonas aeruginosa are central fungal and bacterial members of the pulmonary microbiota. The interactions between A. fumigatus and P. aeruginosa have only just begun to be explored. A balance between inhibitory and stimulatory effects on fungal growth was observed in mixed A. fumigatus–P. aeruginosa cultures. Negative interactions have been seen for homoserine-lactones, pyoverdine and pyochelin resulting from iron starvation and intracellular inhibitory reactive oxidant production. In contrast, several types of positive interactions were recognized. Dirhamnolipids resulted in the production of a thick fungal cell wall, allowing the fungus to resist stress. Phenazines and pyochelin favor iron uptake for the fungus. A. fumigatus is able to use bacterial volatiles to promote its growth. The immune response is also differentially regulated by co-infections. Full article
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Open AccessArticle
Accuracy of Buffy Coat in the Diagnosis of Disseminated Histoplasmosis in AIDS-Patients in an Endemic Area of Brazil
J. Fungi 2019, 5(2), 47; https://doi.org/10.3390/jof5020047
Received: 1 May 2019 / Revised: 28 May 2019 / Accepted: 31 May 2019 / Published: 9 June 2019
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Abstract
The buffy coat is obtained routinely for disseminated histoplamosis (DH) diagnosis in Ceará, Brazil. The aim of this study is to describe the accuracy of staining smears for Histoplasma in the buffy coat of AIDS-patients with DH. From 2012–2013, all results of stained [...] Read more.
The buffy coat is obtained routinely for disseminated histoplamosis (DH) diagnosis in Ceará, Brazil. The aim of this study is to describe the accuracy of staining smears for Histoplasma in the buffy coat of AIDS-patients with DH. From 2012–2013, all results of stained buffy coat smears and culture for fungi performed at São José Hospital were recorded. In total, 489 buffy coats of 361 patients were studied; 19/361 (5.3%; 95%CI = 2.9–7.6%) had positive direct examination stained smears for Histoplasma and 61/361 (16.9%; 95%CI = 13.0–20.8%) had growth in culture. For those with positive Histoplasma cultures, the CD4 count was significantly lower (139.3 vs. 191.7cells/µL; p = 0.014) than others, and death was 18%. The sensitivity and specificity of stained smears was 25.9% and 100%, respectively. A second test, performed up to 36 days from the first one, increased the sensitivity of stained smears to 32.2%. Stained smears of buffy coat have low accuracy; nonetheless, they are easy to perform and can give a quick diagnosis in low-resource endemic areas. Despite the decrease in mortality, it is not yet to the low levels observed in areas that have better and more efficient methods. Full article
(This article belongs to the Special Issue Histoplasma and Histoplasmosis)
Open AccessCommunication
Influence of Environmental Growth Factors on the Biomass and Pigment Production of Chlorociboria aeruginascens
J. Fungi 2019, 5(2), 46; https://doi.org/10.3390/jof5020046
Received: 28 March 2019 / Revised: 24 May 2019 / Accepted: 6 June 2019 / Published: 8 June 2019
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Abstract
The soft rot fungus Chlorociboria aeruginascens produces a blue–green pigment xylindein, which is of considerable interest for various applications such as in the veneer industry or in organic semiconductors. To understand the fungal growth as well as pigment production of C. aeruginascens, [...] Read more.
The soft rot fungus Chlorociboria aeruginascens produces a blue–green pigment xylindein, which is of considerable interest for various applications such as in the veneer industry or in organic semiconductors. To understand the fungal growth as well as pigment production of C. aeruginascens, several studies were performed, the results of which are presented here. These studies investigated various growth conditions such as temperature, pH value, oxygen level and light intensity. It was observed that the formation of xylindein by C. aeruginascens decoupled from growth. In the primary metabolismus, the uncolored biomass is formed. Pigment production took place within the secondary metabolism, while biomass growth as well as pigment production depended on various growth conditions. It was also found that certain conditions encourage the switch in metabolism, leading to pigment production. Full article
(This article belongs to the Special Issue Fungal Polyketides and Other Secondary Metabolites)
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Open AccessReview
Histoplasma Responses to Nutritional Immunity Imposed by Macrophage Activation
J. Fungi 2019, 5(2), 45; https://doi.org/10.3390/jof5020045
Received: 13 May 2019 / Revised: 31 May 2019 / Accepted: 31 May 2019 / Published: 5 June 2019
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Abstract
The fungal pathogen Histoplasma capsulatum resides within the phagosome of host phagocytic cells. Within this intracellular compartment, Histoplasma yeast replication requires the acquisition of several essential nutrients, including metal ions. Recent work has shown that while iron, zinc, and copper are sufficiently abundant [...] Read more.
The fungal pathogen Histoplasma capsulatum resides within the phagosome of host phagocytic cells. Within this intracellular compartment, Histoplasma yeast replication requires the acquisition of several essential nutrients, including metal ions. Recent work has shown that while iron, zinc, and copper are sufficiently abundant in resting macrophages, cytokine activation of these host cells causes restriction of these metals from intracellular yeasts as a form of nutritional immunity. Faced with limited iron availability in the phagosome following macrophage activation by IFN-γ, Histoplasma yeasts secrete iron-scavenging siderophores and employ multiple strategies for reduction of ferric iron to the more physiologically useful ferrous form. IFN-γ activation of macrophages also limits availability of copper in the phagosome, forcing Histoplasma reliance on the high affinity Ctr3 copper importer for copper acquisition. GM-CSF activation stimulates macrophage production of zinc-chelating metallothioneins and zinc transporters to sequester zinc from Histoplasma yeasts. In response, Histoplasma yeasts express the Zrt2 zinc importer. These findings highlight the dynamics of phagosomal metal ion concentrations in host-pathogen interactions and explain one mechanism by which macrophages become a less permissive environment for Histoplasma replication with the onset of adaptive immunity. Full article
(This article belongs to the Special Issue Histoplasma and Histoplasmosis)
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Open AccessArticle
Does Online Search Behavior Coincide with Candida auris Cases? An Exploratory Study
J. Fungi 2019, 5(2), 44; https://doi.org/10.3390/jof5020044
Received: 15 April 2019 / Revised: 16 May 2019 / Accepted: 19 May 2019 / Published: 4 June 2019
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Abstract
Candida auris is an emerging multidrug resistant infectious yeast which is challenging to eradicate and despite available laboratory methods is still difficult to identify especially in less developed countries. To limit the rapid spread of C. auris, quick and accurate detection is [...] Read more.
Candida auris is an emerging multidrug resistant infectious yeast which is challenging to eradicate and despite available laboratory methods is still difficult to identify especially in less developed countries. To limit the rapid spread of C. auris, quick and accurate detection is essential. From the perspective of disease surveillance, additional methods of tracking this yeast are needed. In order to increase global preparedness, we explored the use of online search behavior to monitor the recent global spread of C. auris. We used Google Trends to assess online search behavior on C. auris from January 2016 until August 2018. Weekly Google Trends results were counted as hits and compared to confirmed C. auris cases obtained via publications and a global expert network of key opinion leaders. A total of 44 countries generated a hit, of which 30% (13/44) were confirmed known cases, 34% (15/44) were missed known cases, 34% (15/44) were hits for unknown cases, and 2% (1/44) were confirmed unknown cases. Conclusions: Google Trends searches is rapidly able to provide information on countries with an increased search interest in C. auris. However, Google Trends search results do not generally coincide with C. auris cases or clusters. This study did show that using Google Trends provides both insight into the known and highlights the unknown, providing potential for surveillance and tracking and hence aid in taking timely precautionary measures. Full article
(This article belongs to the Special Issue Candida auris)
Open AccessReview
Endophytic Fungi from Terminalia Species: A Comprehensive Review
J. Fungi 2019, 5(2), 43; https://doi.org/10.3390/jof5020043
Received: 22 April 2019 / Revised: 21 May 2019 / Accepted: 23 May 2019 / Published: 24 May 2019
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Abstract
Endophytic fungi have proven their usefulness for drug discovery, as suggested by the structural complexity and chemical diversity of their secondary metabolites. The diversity and biological activities of endophytic fungi from the Terminalia species have been reported. Therefore, we set out to discuss [...] Read more.
Endophytic fungi have proven their usefulness for drug discovery, as suggested by the structural complexity and chemical diversity of their secondary metabolites. The diversity and biological activities of endophytic fungi from the Terminalia species have been reported. Therefore, we set out to discuss the influence of seasons, locations, and even the plant species on the diversity of endophytic fungi, as well as their biological activities and secondary metabolites isolated from potent strains. Our investigation reveals that among the 200–250 Terminalia species reported, only thirteen species have been studied so far for their endophytic fungi content. Overall, more than 47 fungi genera have been reported from the Terminalia species, and metabolites produced by some of these fungi exhibited diverse biological activities including antimicrobial, antioxidant, antimalarial, anti-inflammatory, anti-hypercholesterolemic, anticancer, and biocontrol varieties. Moreover, more than 40 compounds with eighteen newly described secondary metabolites were reported; among these, metabolites are the well-known anticancer drugs, a group that includes taxol, antioxidant compounds, isopestacin, and pestacin. This summary of data illustrates the considerable diversity and biological potential of fungal endophytes of the Terminalia species and gives insight into important findings while paving the way for future investigations. Full article
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Open AccessArticle
HIV-Associated Cryptococcal Immune Reconstitution Inflammatory Syndrome Is Associated with Aberrant T Cell Function and Increased Cytokine Responses
J. Fungi 2019, 5(2), 42; https://doi.org/10.3390/jof5020042
Received: 5 April 2019 / Revised: 24 April 2019 / Accepted: 29 April 2019 / Published: 23 May 2019
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Abstract
Cryptococcal meningitis remains a significant opportunistic infection among HIV-infected patients, contributing 15–20% of HIV-related mortality. A complication of initiating antiretroviral therapy (ART) following opportunistic infection is immune reconstitution inflammatory syndrome (IRIS). IRIS afflicts 10–30% of HIV-infected patients with cryptococcal meningitis (CM), but its [...] Read more.
Cryptococcal meningitis remains a significant opportunistic infection among HIV-infected patients, contributing 15–20% of HIV-related mortality. A complication of initiating antiretroviral therapy (ART) following opportunistic infection is immune reconstitution inflammatory syndrome (IRIS). IRIS afflicts 10–30% of HIV-infected patients with cryptococcal meningitis (CM), but its immunopathogenesis is poorly understood. We compared circulating T cell memory subsets and cytokine responses among 17 HIV-infected Ugandans with CM: 11 with and 6 without CM-IRIS. At meningitis diagnosis, stimulation with cryptococcal capsule component, glucuronoxylomannan (GXM) elicited consistently lower frequencies of CD4+ and CD8+ T cell memory subsets expressing intracellular cytokines (IL-2, IFN-γ, and IL-17) among subjects who subsequently developed CM-IRIS. After ART initiation, T cells evolved to show a decreased CD8+ central memory phenotype. At the onset of CM-IRIS, stimulation more frequently generated polyfunctional IL-2+/IL-17+ CD4+ T cells in patients with CM-IRIS. Moreover, CD8+ central and effector memory T cells from CM-IRIS subjects also demonstrated more robust IL-2 responses to antigenic stimulation vs. controls. Thus, ART during CM elicits distinct differences in T cell cytokine production in response to cryptococcal antigens both prior to and during the development of IRIS, suggesting an immunologic foundation for the development of this morbid complication of CM infection. Full article
(This article belongs to the Special Issue Fungal Infections of the Central Nervous System)
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Open AccessArticle
Antifungal Resistance in Clinical Isolates of Aspergillus spp.: When Local Epidemiology Breaks the Norm
J. Fungi 2019, 5(2), 41; https://doi.org/10.3390/jof5020041
Received: 14 March 2019 / Revised: 13 May 2019 / Accepted: 16 May 2019 / Published: 21 May 2019
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Abstract
Aspergillosis is a set of very frequent and widely distributed opportunistic diseases. Azoles are the first choice for most clinical forms. However, the distribution of azole-resistant strains is not well known around the world, especially in developing countries. The aim of our study [...] Read more.
Aspergillosis is a set of very frequent and widely distributed opportunistic diseases. Azoles are the first choice for most clinical forms. However, the distribution of azole-resistant strains is not well known around the world, especially in developing countries. The aim of our study was to determine the proportion of non-wild type strains among the clinical isolates of Aspergillus spp. To this end, the minimum inhibitory concentration of three azoles and amphotericin B (used occasionally in severe forms) was studied by broth microdilution. Unexpectedly, it was found that 8.1% of the isolates studied have a diminished susceptibility to itraconazole. This value turned out to be similar to the highest azole resistance rate reported in different countries across the world. Full article
(This article belongs to the Special Issue Fungal Epidemiology)
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Open AccessCommunication
Influence of the Nutrients on the Biomass and Pigment Production of Chlorociboria aeruginascens
J. Fungi 2019, 5(2), 40; https://doi.org/10.3390/jof5020040
Received: 28 March 2019 / Revised: 13 May 2019 / Accepted: 14 May 2019 / Published: 16 May 2019
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Abstract
The blue-green pigment xylindein, produced by the soft rot fungus Chlorociboria aeruginascens, is of considerable interest for various applications such as the veneer industry or organic semiconductors. The studies presented were performed in order to understand the fungal growth as well as [...] Read more.
The blue-green pigment xylindein, produced by the soft rot fungus Chlorociboria aeruginascens, is of considerable interest for various applications such as the veneer industry or organic semiconductors. The studies presented were performed in order to understand the fungal growth as well as the pigment production of C. aeruginascens. Therefore, various nutrient compositions were investigated. As a result, observations of the formation of xylindein through C. aeruginascens decoupling from growth were made. In the primary metabolism the uncolored biomass is formed. Various carbohydrates were determined as nutrients for the fungus and as a nitrogen source it was observed that the fungus prefers the complex organic nitrogen source, that being yeast extract. Furthermore, it was discovered that the ratio between carbohydrate and nitrogen sources encourages the switch of the metabolism and therewith the production of the blue-green pigment xylindein. Full article
(This article belongs to the Special Issue Fungal Polyketides and Other Secondary Metabolites)
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Open AccessArticle
A Universally Primed-Polymerase Chain Reaction (UP-PCR) Marker to Discriminate Clonostachys rosea ACM941 from Related Strains
J. Fungi 2019, 5(2), 39; https://doi.org/10.3390/jof5020039
Received: 2 April 2019 / Revised: 2 May 2019 / Accepted: 13 May 2019 / Published: 14 May 2019
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Abstract
Clonostachys rosea strain ACM941 is an effective biocontrol agent against several crop diseases including Fusarium head blight. In anticipation of its increased relevance going forward, the development of a reliable DNA-based molecular marker to track it is essential. Universally primed-PCR (UP-PCR) has been [...] Read more.
Clonostachys rosea strain ACM941 is an effective biocontrol agent against several crop diseases including Fusarium head blight. In anticipation of its increased relevance going forward, the development of a reliable DNA-based molecular marker to track it is essential. Universally primed-PCR (UP-PCR) has been used successfully to differentiate other C. rosea strains. Herein, the development of a UP-PCR marker for ACM941 is described. A combination of two primers (AS15 and L45) produced a ~450 bp fragment that was unique to ACM941 compared to other commercial biocontrol agents. Primers subsequently designed based on the obtained fragment also produced a similarly unique band from ACM941 alone. BLAST analysis of the amplified sequence did not yield any homologous sequence in available online databases or within the closely related C. rosea IK726 and CBS125111 strains’ genomes. The specificity of this marker for ACM941 was validated against ten additional C. rosea strains isolated from Canada, with ACM941 producing the brightest band. Taken together, these results imply that the UP-PCR primers AS15 and L45 and the amplified fragment can be used to detect and monitor the ACM941 strain after its release into the environment. Full article
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Open AccessArticle
Estimated Burden of Serious Fungal Infections in Ghana
J. Fungi 2019, 5(2), 38; https://doi.org/10.3390/jof5020038
Received: 5 March 2019 / Revised: 11 April 2019 / Accepted: 14 April 2019 / Published: 11 May 2019
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Abstract
Fungal infections are increasingly becoming common and yet often neglected in developing countries. Information on the burden of these infections is important for improved patient outcomes. The burden of serious fungal infections in Ghana is unknown. We aimed to estimate this burden. Using [...] Read more.
Fungal infections are increasingly becoming common and yet often neglected in developing countries. Information on the burden of these infections is important for improved patient outcomes. The burden of serious fungal infections in Ghana is unknown. We aimed to estimate this burden. Using local, regional, or global data and estimates of population and at-risk groups, deterministic modelling was employed to estimate national incidence or prevalence. Our study revealed that about 4% of Ghanaians suffer from serious fungal infections yearly, with over 35,000 affected by life-threatening invasive fungal infections. Incidence of cryptococcal meningitis, Pneumocystis jirovecii pneumonia, and disseminated histoplasmosis cases in AIDS was estimated at 6275, 12,610 and 724, respectively. Oral and esophageal candidiasis collectively affect 27,100 Ghanaians and 42,653 adult asthmatics are estimated to have fungal asthma. We estimate a prevalence of 12,620 cases of chronic pulmonary aspergillosis (CPA and an incidence of 1254 cases of invasive aspergillosis (IA). Estimated cases of candidemia and candida peritonitis cases were 1446 and 217, respectively. The estimated prevalence of recurrent vulvovaginal candidiasis (RVVC) and tinea capitis was 442,621 and 598,840, respectively. Mucormycosis and fungal keratitis each may affect 58 and 810 Ghanaians. These data highlight the urgent need for intensified awareness to improve diagnosis and management. Full article
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Open AccessReview
Candida/Staphylococcal Polymicrobial Intra-Abdominal Infection: Pathogenesis and Perspectives for a Novel Form of Trained Innate Immunity
J. Fungi 2019, 5(2), 37; https://doi.org/10.3390/jof5020037
Received: 15 April 2019 / Revised: 6 May 2019 / Accepted: 7 May 2019 / Published: 9 May 2019
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Abstract
Polymicrobial sepsis is difficult to diagnose and treat and causes significant morbidity and mortality, especially when fungi are involved. In vitro, synergism between Candida albicans and various bacterial species has been described for many years. Our laboratory has developed a murine model of [...] Read more.
Polymicrobial sepsis is difficult to diagnose and treat and causes significant morbidity and mortality, especially when fungi are involved. In vitro, synergism between Candida albicans and various bacterial species has been described for many years. Our laboratory has developed a murine model of polymicrobial intra-abdominal infection with Candida albicans and Staphylococcus aureus, demonstrating that polymicrobial infections cause high levels of mortality, while monoinfections do not. By contrast, closely related Candida dubliniensis does not cause synergistic lethality and rather provides protection against lethal polymicrobial infection. This protection is thought to be driven by a novel form of trained innate immunity mediated by myeloid-derived suppressor cells (MDSCs), which we are proposing to call “trained tolerogenic immunity”. MDSC accumulation has been described in patients with sepsis, as well as in in vivo sepsis models. However, clinically, MDSCs are considered detrimental in sepsis, while their role in in vivo models differs depending on the sepsis model and timing. In this review, we will discuss the role of MDSCs in sepsis and infection and summarize our perspectives on their development and function in the spectrum of trained innate immune protection against fungal-bacterial sepsis. Full article
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Open AccessArticle
An Ecological Assessment of Isaria fumosorosea Applications Compared to a Neonicotinoid Treatment for Regulating Invasive Ficus Whitefly
J. Fungi 2019, 5(2), 36; https://doi.org/10.3390/jof5020036
Received: 1 March 2019 / Revised: 24 April 2019 / Accepted: 25 April 2019 / Published: 4 May 2019
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Abstract
A pilot study was conducted on a weeping fig, Ficus benjamina shrub hedge in a Florida urban landscape to determine the efficacy of a fungal biopesticide, PFR-97™ which contains blastospores of Isaria fumosorosea, and a neonicotinoid treatment (Admire Pro™) applied against the [...] Read more.
A pilot study was conducted on a weeping fig, Ficus benjamina shrub hedge in a Florida urban landscape to determine the efficacy of a fungal biopesticide, PFR-97™ which contains blastospores of Isaria fumosorosea, and a neonicotinoid treatment (Admire Pro™) applied against the invasive ficus whitefly pest, Singhiella simplex (Singh). Post treatment, an ecological assessment of the study was conducted by observing the impact of the fungal biopesticide and neonicotinoid treatment on natural enemies, e.g., predators, parasitoids and enzootic fungal pathogens occurring in the whitefly-infested hedge. Both treatments provided a significant reduction in the whitefly population compared to control and were compatible with the natural enemies present. Various natural enemies including fungal entomopathogens were identified associated with the whitefly population infesting the weeping fig hedge. The parasitoids, Encarsia protransvena Viggiani and Amitus bennetti Viggiani & Evans combined parasitized a similar mean number of whitefly nymphs in both treatments and control; however, the number parasitized decreased over time. Natural enzootic fungi isolated from the ficus whitefly nymphs were I. fumosorosea, Purpureocillium lilacinum and Lecanicillium, Aspergillus and Fusarium species. Results from this pilot study suggest there is much potential for using repeated applications of the fungal biopesticide, PFR-97™ as a foliar spray compared to a neonicitionid as a soil drench for managing S. simplex on Ficus species for ≥28 days. Full article
(This article belongs to the Special Issue Fungal-Insect Interactions)
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Open AccessArticle
Epidemiological and Mycological Aspects of Onychomycosis in Dakar (Senegal)
J. Fungi 2019, 5(2), 35; https://doi.org/10.3390/jof5020035
Received: 15 March 2019 / Revised: 16 April 2019 / Accepted: 24 April 2019 / Published: 29 April 2019
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Abstract
Onychomycosis is a fungal nails infection often caused by yeasts, dermatophytes and molds. It is an important public health concern due to its high prevalence, the problem of diagnostics, and the poor response to treatments. The objective of this study was to evaluate [...] Read more.
Onychomycosis is a fungal nails infection often caused by yeasts, dermatophytes and molds. It is an important public health concern due to its high prevalence, the problem of diagnostics, and the poor response to treatments. The objective of this study was to evaluate the epidemiological and microbiological profile of onychomycosis diagnosed at the Laboratory of Parasitology-Mycology of the National University Hospital of Fann in Dakar, Senegal, from 2012 to 2016. A retrospective and descriptive study was performed from January 2012 to December 2016 in a patient attending the laboratory of Parasitology-Mycology at the Fann teaching hospital. Socio-demographic, clinical and biological data were collected from the bench registers. Samples from the lesions were tested using direct microscopy and cultured on a Sabouraud-Chloramphenicol and Sabouraud-Chloramphenicol-Actidione medium. A descriptive analysis was done using Stata IC 12 software. The significance level of different tests was set at 5% two-side. A total of 469 patients were included in this study. The mean age of the study population was 33.2 ± 15.2 years, and the sex ratio was 0.52. The prevalence of onychomycosis was 48.4% (227/469). The main clinical presentations were disto-lateral subungual onychomycosis (37.9%) and onyxis (46.5%). Identified fungal species were Candida albicans (42.7%), Candida spp. (39.5%), Trichophyton soudanense (10.1%), Fusarium spp. (5.3%), and Candida tropicalis (2.6%). Candida albicans was more frequent in subjects over 15 years of age (43.6%) and women (45%). However, Trichophyton soudanense was higher in patients under 15 years old (17.4%) as well as in male subjects (18.8%). In conclusion, onychomycosis is a common cause of consultation in health facilities. Candida albicans and Trichophyton Soudanense are the main fungal species causing onychomycosis. A better understanding of the epidemiology of onychomycosis as well as the spectrum of the pathogen could contribute to improve the management of the infection. Full article
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Open AccessReview
Beyond Antagonism: The Interaction Between Candida Species and Pseudomonas aeruginosa
J. Fungi 2019, 5(2), 34; https://doi.org/10.3390/jof5020034
Received: 19 March 2019 / Revised: 18 April 2019 / Accepted: 18 April 2019 / Published: 19 April 2019
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Abstract
There are many examples of the interaction between prokaryotes and eukaryotes. One such example is the polymicrobial colonization/infection by the various opportunistic pathogenic yeasts belonging to the genus Candida and the ubiquitous bacterium, Pseudomonas aeruginosa. Although this interaction has simplistically been characterized [...] Read more.
There are many examples of the interaction between prokaryotes and eukaryotes. One such example is the polymicrobial colonization/infection by the various opportunistic pathogenic yeasts belonging to the genus Candida and the ubiquitous bacterium, Pseudomonas aeruginosa. Although this interaction has simplistically been characterized as antagonistic to the yeast, this review highlights the complexity of the interaction with various factors influencing both microbes. The first section deals with the interactions in vitro, looking specifically at the role of cell wall components, quorum sensing molecules, phenazines, fatty acid metabolites and competition for iron in the interaction. The second part of this review places all these interactions in the context of various infection or colonization sites, i.e., lungs, wounds, and the gastrointestinal tract. Here we see that the role of the host, as well as the methodology used to establish co-infection, are important factors, influencing the outcome of the disease. Suggested future perspectives for the study of this interaction include determining the influence of newly identified participants of the QS network of P. aeruginosa, oxylipin production by both species, as well as the genetic and phenotypic plasticity of these microbes, on the interaction and outcome of co-infection. Full article
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Open AccessReview
Is the Insect Cuticle the only Entry Gate for Fungal Infection? Insights into Alternative Modes of Action of Entomopathogenic Fungi
J. Fungi 2019, 5(2), 33; https://doi.org/10.3390/jof5020033
Received: 28 February 2019 / Revised: 11 April 2019 / Accepted: 12 April 2019 / Published: 16 April 2019
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Abstract
Entomopathogenic fungi are the only insect pathogens able to infect their host by adhesion to the surface and penetration through the cuticle. Although the possibility of fungal infection per os was described almost a century ago, there is an information gap of several [...] Read more.
Entomopathogenic fungi are the only insect pathogens able to infect their host by adhesion to the surface and penetration through the cuticle. Although the possibility of fungal infection per os was described almost a century ago, there is an information gap of several decades regarding this topic, which was poorly explored due to the continuous elucidation of cuticular infection processes that lead to insect death by mycosis. Recently, with the advent of next-generation sequencing technologies, the genomes of the main entomopathogenic fungi became available, and many fungal genes potentially useful for oral infection were described. Among the entomopathogenic Hypocreales that have been sequenced, Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Cordycipitaceae) is the main candidate to explore this pathway since it has a major number of shared genes with other non-fungal pathogens that infect orally, such as Bacillus thuringiensis Berliner (Bacillales: Bacillaceae). This finding gives B. bassiana a potential advantage over other entomopathogenic fungi: the possibility to infect through both routes, oral and cuticular. In this review, we explore all known entry gates for entomopathogenic fungi, with emphasis on the infection per os. We also set out the fungal infection process in a more integral approach, as a need to exploit its full potential for insect control, considering all of its virulence factors and the conditions needed to improve its virulence against insect that might offer some resistance to the common infection through the cuticle. Full article
(This article belongs to the Special Issue Fungal-Insect Interactions)
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Open AccessArticle
Clonal Dispersal of Cryptococcus gattii VGII in an Endemic Region of Cryptococcosis in Colombia
J. Fungi 2019, 5(2), 32; https://doi.org/10.3390/jof5020032
Received: 8 March 2019 / Revised: 4 April 2019 / Accepted: 6 April 2019 / Published: 15 April 2019
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Abstract
This study characterized the genotype and phenotype of Cryptococcus gattii VGII isolates from Cucuta, an endemic region of cryptococcal disease in Colombia, and compared these traits with those from representative isolates from the Vancouver Island outbreak (VGIIa and VGIIb). Genetic diversity was assessed [...] Read more.
This study characterized the genotype and phenotype of Cryptococcus gattii VGII isolates from Cucuta, an endemic region of cryptococcal disease in Colombia, and compared these traits with those from representative isolates from the Vancouver Island outbreak (VGIIa and VGIIb). Genetic diversity was assessed by multilocus sequence typing (MLST) analysis. Phenotypic characteristics, including growth capacity under different temperature and humidity conditions, macroscopic and microscopic morphology, phenotypic switching, mating type, and activity of extracellular enzymes were studied. Virulence was studied in vivo in a mouse model. MLST analysis showed that the isolates from Cucuta were highly clonal, with ST25 being the most common genotype. Phenotypically, isolates from Cucuta showed large cell and capsular sizes, and shared phenotypic traits and enzymatic activities among them. The mating type a prevailed among the isolates, which were fertile and of considerable virulence in the animal model. This study highlights the need for a continuous surveillance of C. gattii in Colombia, especially in endemic areas like Cucuta, where the highest number of cryptococcosis cases due to this species is reported. This will allow the early detection of potentially highly virulent strains that spread clonally, and can help prevent the occurrence of outbreaks in Colombia and elsewhere. Full article
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Open AccessReview
The Spectrum of Interactions between Cryptococcus neoformans and Bacteria
J. Fungi 2019, 5(2), 31; https://doi.org/10.3390/jof5020031
Received: 27 March 2019 / Revised: 8 April 2019 / Accepted: 11 April 2019 / Published: 12 April 2019
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Abstract
Cryptococcus neoformans is a major fungal pathogen that infects immunocompromised people and causes life-threatening meningoencephalitis. C. neoformans does not occur in isolation either in the environment or in the human host, but is surrounded by other microorganisms. Bacteria are ubiquitously distributed in nature, [...] Read more.
Cryptococcus neoformans is a major fungal pathogen that infects immunocompromised people and causes life-threatening meningoencephalitis. C. neoformans does not occur in isolation either in the environment or in the human host, but is surrounded by other microorganisms. Bacteria are ubiquitously distributed in nature, including soil, and make up the dominant part of the human microbiota. Pioneering studies in the 1950s demonstrated antifungal activity of environmental bacteria against C. neoformans. However, the mechanisms and implications of these interactions remain largely unknown. Recently, interest in polymicrobial interaction studies has been reignited by the development of improved sequencing methodologies, and by the realization that such interactions may have a huge impact on ecology and human health. In this review, we summarize our current understanding of the interaction of bacteria with C. neoformans. Full article
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Open AccessArticle
Yeast Species in the Oral Cavities of Older People: A Comparison between People Living in Their Own Homes and Those in Rest Homes
J. Fungi 2019, 5(2), 30; https://doi.org/10.3390/jof5020030
Received: 26 February 2019 / Revised: 9 April 2019 / Accepted: 9 April 2019 / Published: 12 April 2019
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Abstract
Oral candidiasis is prevalent among older people due to predisposing factors such as impaired immune defenses, medications and denture use. An increasing number of older people live in rest home facilities and it is unclear how this institutionalized living affects the quantity and [...] Read more.
Oral candidiasis is prevalent among older people due to predisposing factors such as impaired immune defenses, medications and denture use. An increasing number of older people live in rest home facilities and it is unclear how this institutionalized living affects the quantity and type of fungi colonizing these people’s oral cavities. Smears and swabs of the palate and tongue and saliva samples were taken from participants residing in rest homes (RH; n = 20) and older people living in their own homes (OH; n = 20). Yeast in samples were quantified and identified by culturing on CHROMagar Candida and sequencing the ITS2 region of rDNA. A higher proportion of RH residents had Candida hyphae present in smears compared to OH participants (35% vs. 30%) although this difference was not statistically significant (p = 0.74). RH residents had, on average, 23 times as many yeast per mL saliva as OH participants (p = 0.01). Seven yeast species were identified in OH samples and only five in RH samples, with Candida albicans and Candida glabrata being the most common species isolated from both participant groups. The results indicate that older people living in aged-care facilities were more likely to have candidiasis and have a higher yeast carriage rate than similarly aged people living at home. This may be due to morbidities which led to the need for residential care and/or related to the rest home environment. Full article
(This article belongs to the Special Issue Oral Mycobiome)
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Open AccessReview
A Bright Future for Fluorescence Imaging of Fungi in Living Hosts
J. Fungi 2019, 5(2), 29; https://doi.org/10.3390/jof5020029
Received: 25 February 2019 / Revised: 27 March 2019 / Accepted: 29 March 2019 / Published: 3 April 2019
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Abstract
Traditional in vivo investigation of fungal infection and new antifungal therapies in mouse models is usually carried out using post mortem methodologies. However, biomedical imaging techniques focusing on non-invasive techniques using bioluminescent and fluorescent proteins have become valuable tools. These new techniques address [...] Read more.
Traditional in vivo investigation of fungal infection and new antifungal therapies in mouse models is usually carried out using post mortem methodologies. However, biomedical imaging techniques focusing on non-invasive techniques using bioluminescent and fluorescent proteins have become valuable tools. These new techniques address ethical concerns as they allow reduction in the number of animals required to evaluate new antifungal therapies. They also allow better understanding of the growth and spread of the pathogen during infection. In this review, we concentrate on imaging technologies using different fungal reporter proteins. We discuss the advantages and limitations of these different reporters and compare the efficacy of bioluminescent and fluorescent proteins for fungal research. Full article
Open AccessArticle
Candida albicans and non-albicans Isolates from Bloodstream Have Different Capacities to Induce Neutrophil Extracellular Traps
J. Fungi 2019, 5(2), 28; https://doi.org/10.3390/jof5020028
Received: 11 March 2019 / Revised: 27 March 2019 / Accepted: 28 March 2019 / Published: 1 April 2019
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Abstract
Neutrophils activated with pathogens or their products induce formation of extracellular traps (NETs), but if this constitutes a general response against all pathogenic species in a single genus or intrageneric differences exist remains unknown, yet this is of great importance for the establishment [...] Read more.
Neutrophils activated with pathogens or their products induce formation of extracellular traps (NETs), but if this constitutes a general response against all pathogenic species in a single genus or intrageneric differences exist remains unknown, yet this is of great importance for the establishment of effective treatments. To determine this, we analyzed neutrophil extracellular traps formation after the stimulation with bloodstream isolates from different Candida species (Candida albicans, C. tropicalis, C. parapsilosis, and C. glabrata), and found that each species has a different capacity to induce DNA extrusion, which is independent of their morphology (yeast or hyphae). We observed that phospholipase producer’s strains and their secretion products were able to induce NETs, a property not observed with phospholipase deficient strains, with exception of some Candida glabrata sensu stricto isolates, which showed no NETs induction although they did show phospholipase production. To further analyze this, we extended our study to include Candida glabrata cryptic species (C. bracarensis and C. nivariensis) and no extracellular traps formation was observed. Here, we contribute to the understanding of how neutrophils initiate NETs, and we found that certain strains may have a differential capacity to trigger these structures, which may explain the high mortality of some isolates. Full article
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Open AccessReview
Animal Models to Study Mucormycosis
J. Fungi 2019, 5(2), 27; https://doi.org/10.3390/jof5020027
Received: 21 February 2019 / Revised: 22 March 2019 / Accepted: 26 March 2019 / Published: 27 March 2019
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Abstract
Mucormycosis is a rare but often fatal or debilitating infection caused by a diverse group of fungi. Animal models have been crucial in advancing our knowledge of mechanisms influencing the pathogenesis of mucormycoses, and to evaluate therapeutic strategies. This review describes the animal [...] Read more.
Mucormycosis is a rare but often fatal or debilitating infection caused by a diverse group of fungi. Animal models have been crucial in advancing our knowledge of mechanisms influencing the pathogenesis of mucormycoses, and to evaluate therapeutic strategies. This review describes the animal models established for mucormycosis, summarizes how they have been applied to study mucormycoses, and discusses the advantages and limitations of the different model systems. Full article
(This article belongs to the Special Issue Mucorales and Mucormycosis)
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