Abstract:This supplement of 'Mycoses' is devoted to infections caused by a group of fungi traditionally known as the zygomycetes. The Zygomycota represent an important group of medically important opportunistic fungi, which cause devastating fungal infections in humans and animals with severe underlying immune or metabolic disorders. These infections are increasing in numbers due to the growing populations of patients with uncontrolled diabetes and immunosuppression, as well as the increased use of prophylactic measure… Show more
“…However, we recently showed that the early stages of infection control are critical for predicting disease outcome 41 . Consistent with this, the majority of mucormycosis patients have predisposing factors that impact phagocyte recruitment, including iron overload, diabetes mellitus, neutropenia, organ transplantation, trauma, and corticosteroid therapy 19,22,34,42-50 . Outbreaks of R. microsporus in hospitals have been linked to the use of contaminated supplies during treatment of immunocompromised patients 51-55 .…”
Section: Introductionmentioning
confidence: 56%
“…Pathogenic Mucorales span multiple genera, with the Rhizopus genus causing almost half of all documented cases and high mortality in susceptible patient populations (5)(6)(7)(8)(9)(10)(11)(12).…”
Environmentally ubiquitous fungal spores of the Mucorales order cause acute invasive infections through germination and evasion of the mammalian host immune system. Early phagocyte control of spore germination plays a key role in controlling infection, yet swelling Mucorales spores evade phagocytosis through an unknown mechanism. Here we investigate fungal immune evasion in a clinical isolate of Rhizopus microsporus and reveal the role of a bacterial endosymbiont, Ralsonia pickettii, in fungal pathogenesis. Analysis of phagocytosis rates in wild type and cured fungal isolates demonstrates a role for the endosymbiont in immune cell evasion through disruption of the cytoskeleton and phagosome maturation. Further analysis of bacterial secreted products revealed the presence of a previously uncharacterized secondary metabolite whose production is induced by the presence of the fungus. Analysis of the bacterial genome and condition-dependent RNAseq implicate a cryptic type I polyketide synthase. Subsequent analysis of wild type and cured spores in a zebrafish larvae model of infection demonstrate a role for the endosymbiont in suppressing macrophage and neutrophil recruitment to the site of infection. Finally, we demonstrate that this has implications for fungal clearance in an immunocompetent murine model of infection. Together, these findings identify for the first time a role for a bacterial endosymbiont in the pathogenesis of Rhizopus microsporus during animal infection.
“…However, we recently showed that the early stages of infection control are critical for predicting disease outcome 41 . Consistent with this, the majority of mucormycosis patients have predisposing factors that impact phagocyte recruitment, including iron overload, diabetes mellitus, neutropenia, organ transplantation, trauma, and corticosteroid therapy 19,22,34,42-50 . Outbreaks of R. microsporus in hospitals have been linked to the use of contaminated supplies during treatment of immunocompromised patients 51-55 .…”
Section: Introductionmentioning
confidence: 56%
“…Pathogenic Mucorales span multiple genera, with the Rhizopus genus causing almost half of all documented cases and high mortality in susceptible patient populations (5)(6)(7)(8)(9)(10)(11)(12).…”
Environmentally ubiquitous fungal spores of the Mucorales order cause acute invasive infections through germination and evasion of the mammalian host immune system. Early phagocyte control of spore germination plays a key role in controlling infection, yet swelling Mucorales spores evade phagocytosis through an unknown mechanism. Here we investigate fungal immune evasion in a clinical isolate of Rhizopus microsporus and reveal the role of a bacterial endosymbiont, Ralsonia pickettii, in fungal pathogenesis. Analysis of phagocytosis rates in wild type and cured fungal isolates demonstrates a role for the endosymbiont in immune cell evasion through disruption of the cytoskeleton and phagosome maturation. Further analysis of bacterial secreted products revealed the presence of a previously uncharacterized secondary metabolite whose production is induced by the presence of the fungus. Analysis of the bacterial genome and condition-dependent RNAseq implicate a cryptic type I polyketide synthase. Subsequent analysis of wild type and cured spores in a zebrafish larvae model of infection demonstrate a role for the endosymbiont in suppressing macrophage and neutrophil recruitment to the site of infection. Finally, we demonstrate that this has implications for fungal clearance in an immunocompetent murine model of infection. Together, these findings identify for the first time a role for a bacterial endosymbiont in the pathogenesis of Rhizopus microsporus during animal infection.
“…Pathogenic Mucorales span multiple genera, with the Rhizopus genus causing almost half of all documented cases and high mortality in susceptible patient populations. [6][7][8][9][10][11][12][13] Infection occurs through inoculation with dormant, immunologically inert spores. Upon germination, these spores become metabolically active and begin to swell.…”
A bacterial endosymbiont of the fungus Rhizopus microsporus drives phagocyte evasion and opportunistic virulence Highlights d Bacterial endosymbionts protect fungal spores from phagocytes d A secreted factor blocks growth and killing by environmental amoebas d Endosymbionts improve fungal stress resistance d Endosymbiosis also allows the evasion of vertebrate immune cells and virulence in vivo
“…The incidence of mucormycosis is relatively low, but it constitutes a significant area of concern in immunocompromised patients, including pediatric patients with malignances [ 4 , 5 ]. One of the major problems concerning mucormycosis is the early accurate diagnosis and the identification of the involved organism [ 6 , 7 ].…”
Objectives
The main aim of this work was to compare the methods of DNA isolation in the moulds of genus
Mucorales
with special regard to the amount and purity of the DNA acquired. The acquired DNA was then amplified by specific real-time PCR.
Design
Five DNA extraction procedures were carried out in a Class 2 Biosafety cabinet in a dedicated room with suitable biosafety precautions and appropriate biowaste disposal methods. A total of 6
Mucorales
clinical strains were used.
Results
From the viewpoint of concentration and purity, methods A shown abundant amount of fungal DNA whereas methods E report a pure fungal DNA with R260/280 of 1.7 near the optimal 1.8. The DNA quantity reach statistically difference at ANOVA test with p value 0.0005
Conclusion
Overall, the E method was the most efficient method in the extraction of DNA from fungal cultures compared to the other methods considering time, cost, technical expertise, and instrumentation. Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays
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