scholarly journals Azole resistance mechanisms in pathogenic M. furfur

2021 ◽  
Author(s):  
Cheryl Leong ◽  
Joel Chan Wai Kit ◽  
Shi Mun Lee ◽  
Yuen In Lam ◽  
Joleen P.Z. Goh ◽  
...  
Keyword(s):  
Fungal Pathogens ◽  
Systemic Disease ◽  
Antifungal Susceptibility ◽  
Treatment Strategies ◽  
Systemic Infection ◽  
Resistance Mechanisms ◽  
Fungal Species ◽  
Azole Resistance ◽  
In The Wild

Malassezia are emerging fungal pathogens causing opportunistic skin and severe systemic infection. Nosocomial outbreaks are associated with azole resistance and understanding of the underlying mechanisms are limited to knowledge from other fungal species. Herein, we identified distinct antifungal susceptibility patterns in 26 Malassezia furfur isolates derived from healthy and diseased individuals. A Y67F CYP51 mutation was identified in five isolates of M. furfur. However, this mutation alone was insufficient to induce reduce azole susceptibility in the wild type strain. RNA-seq and differential gene analysis of healthy and disease derived strains exposed to clotrimazole in vitro identified several key metabolic pathways and transporter proteins which are involved in reduce azole susceptibility. The pleiotropic drug transporter PDR10 was the single most highly upregulated transporter gene in multiple strains of M. furfur after azole treatment and increased expression of PDR10 is associated with reduced azole susceptibility in some systemic disease isolates of M. furfur. Deletion of PDR10 in a pathogenic M. furfur strain with reduced susceptibility reduced MIC values to the level of that in susceptible isolates. The current dearth of antifungal technologies, globally emerging multi-azole resistance, and broad agriculture and consumer care use of azoles means improved understanding of the mechanisms underlying intrinsic and acquired azole resistance in Malassezia is crucial for development of antibiotic stewardship and antifungal treatment strategies.

scholarly journals Azole Resistance in Clinical and Environmental Aspergillus Isolates from the French West Indies (Martinique)

2021 ◽  
Vol 7 (5) ◽  
pp. 355
Author(s):  
Lorra Monpierre ◽  
Nicole Desbois-Nogard ◽  
Isabel Valsecchi ◽  
Marielle Bajal ◽  
Cécile Angebault ◽  
...  
Keyword(s):  
West Indies ◽  
Culture Media ◽  
Antifungal Susceptibility ◽  
Resistance Mechanisms ◽  
Clinical Samples ◽  
Azole Resistance ◽  
Environmental Isolates ◽  
French West Indies ◽  
First Time ◽  
Microdilution Broth

The emergence of azole resistant Aspergillus spp., especially Aspergillus fumigatus, has been described in several countries around the world with varying prevalence depending on the country. To our knowledge, azole resistance in Aspergillus spp. has not been reported in the West Indies yet. In this study, we investigated the antifungal susceptibility of clinical and environmental isolates of Aspergillus spp. from Martinique, and the potential resistance mechanisms associated with mutations in cyp51A gene. Overall, 208 Aspergillus isolates were recovered from clinical samples (n = 45) and environmental soil samples (n = 163). They were screened for resistance to azole drugs using selective culture media. The Minimum Inhibitory Concentrations (MIC) towards voriconazole, itraconazole, posaconazole and isavuconazole, as shown by the resistant isolates, were determined using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) microdilution broth method. Eight isolates (A. fumigatus, n = 6 and A. terreus, n = 2) had high MIC for at least one azole drug. The sequencing of cyp51A gene revealed the mutations G54R and TR34/L98H in two A. fumigatus clinical isolates. Our study showed for the first time the presence of azole resistance in A. fumigatus and A. terreus isolates in the French West Indies.

scholarly journals Tamm-Horsfall Protein Protects the Urinary Tract againstCandida albicans

2018 ◽  
Vol 86 (12) ◽  
Author(s):  
Alison Coady ◽  
Anissa R. Ramos ◽  
Joshua Olson ◽  
Victor Nizet ◽  
Kathryn A. Patras
Keyword(s):  
Urinary Tract ◽  
Fungal Pathogens ◽  
Treatment Strategies ◽  
Immune Defense ◽  
Renal Physiology ◽  
Bladder Epithelium ◽  
Indwelling Catheters ◽  
Content Type ◽  
Tract Infections

ABSTRACTUrinary tract infections (UTIs) caused by the human fungal pathogenCandida albicansand related species are prevalent in hospitalized patients, especially those on antibiotic therapy, with indwelling catheters, or with predisposing conditions such as diabetes or immunodeficiency. Understanding of key host defenses againstCandidaUTI is critical for developing effective treatment strategies. Tamm-Horsfall glycoprotein (THP) is the most abundant urine protein, with multiple roles in renal physiology and bladder protection. THP protects against bacterial UTI by blocking bacterial adherence to the bladder epithelium, but its role in defense against fungal pathogens is not yet described. Here we demonstrate that THP restricts colonization of the urinary tract byC. albicans. THP binds toC. albicanshyphae, but not the yeast form, in a manner dependent on fungal expression of the Als3 adhesion glycoprotein. THP directly blocksC. albicansadherence to bladder epithelial cellsin vitro, and THP-deficient mice display increased fungal burden in aC. albicansUTI model. This work outlines a previously unknown role for THP as an essential component for host immune defense against fungal urinary tract infection.

Identification and in Vitro Susceptibility Pattern of Fungal Pathogens in Immunocompromised Patients with invasive Fungal Infections

2021 ◽  
Vol 30 (3) ◽  
pp. 127-134
Author(s):  
Shaimaa A.S. Selem ◽  
Neveen A. Hassan ◽  
Mohamed Z. Abd El-Rahman ◽  
Doaa M. Abd El-Kareem
Keyword(s):  
Intensive Care ◽  
Fungal Infection ◽  
Fungal Pathogens ◽  
Fungal Infections ◽  
Antifungal Susceptibility ◽  
Invasive Fungal Infections ◽  
Immunocompromised Patients ◽  
University Hospitals ◽  
Vitek 2

Background: In intensive care units, invasive fungal infections have become more common, particularly among immunocompromised patients. Early identification and starting the treatment of those patients with antifungal therapy is critical for preventing unnecessary use of toxic antifungal agents. Objective: The aim of this research is to determine which common fungi cause invasive fungal infection in immunocompromised patients, as well as their antifungal susceptibility patterns in vitro, in Assiut University Hospitals. Methodology: This was a hospital based descriptive study conducted on 120 patients with clinical suspicion of having fungal infections admitted at different Intensive Care Units (ICUs) at Assiut University Hospitals. Direct microscopic examination and inoculation on Sabouraud Dextrose Agar (SDA) were performed on the collected specimens. Isolated yeasts were classified using phenotypic methods such as chromogenic media (Brilliance Candida agar), germ tube examination, and the Vitek 2 system for certain isolates, while the identification of mould isolates was primarily based on macroscopic and microscopic characteristics. Moulds were tested in vitro for antifungal susceptibility using the disc diffusion, and yeast were tested using Vitek 2 device cards. Results: In this study, 100 out of 120 (83.3%) of the samples were positive for fungal infection. Candida and Aspergillus species were the most commonly isolated fungal pathogens. The isolates had the highest sensitivity to Amphotericin B (95 %), followed by Micafungin (94 %) in an in vitro sensitivity survey. Conclusion: Invasive fungal infections are a leading cause of morbidity and mortality in immunocompromised patients, with Candida albicans being the most frequently isolated yeast from various clinical specimens; however, the rise in resistance, especially to azoles, is a major concern.

scholarly journals QseC Mediates Salmonella enterica Serovar Typhimurium Virulence In Vitro and In Vivo

2009 ◽  
Vol 78 (3) ◽  
pp. 914-926 ◽  
Author(s):  
Cristiano G. Moreira ◽  
David Weinshenker ◽  
Vanessa Sperandio
Keyword(s):  
Epithelial Cells ◽  
Salmonella Enterica ◽  
Systemic Disease ◽  
Systemic Infection ◽  
Flagellar Motility ◽  
Signaling System ◽  
Interkingdom Signaling ◽  
Serovar Typhimurium

ABSTRACT The autoinducer-3 (AI-3)/epinephrine (Epi)/norepinephrine (NE) interkingdom signaling system mediates chemical communication between bacteria and their mammalian hosts. The three signals are sensed by the QseC histidine kinase (HK) sensor. Salmonella enterica serovar Typhimurium is a pathogen that uses HKs to sense its environment and regulate virulence. Salmonella serovar Typhimurium invades epithelial cells and survives within macrophages. Invasion of epithelial cells is mediated by the type III secretion system (T3SS) encoded in Salmonella pathogenicity island 1 (SPI-1), while macrophage survival and systemic disease are mediated by the T3SS encoded in SPI-2. Here we show that QseC plays an important role in Salmonella serovar Typhimurium pathogenicity. A qseC mutant was impaired in flagellar motility, in invasion of epithelial cells, and in survival within macrophages and was attenuated for systemic infection in 129x1/SvJ mice. QseC acts globally, regulating expression of genes within SPI-1 and SPI-2 in vitro and in vivo (during infection of mice). Additionally, dopamine β-hydroxylase knockout (Dbh − / −) mice that do not produce Epi or NE showed different susceptibility to Salmonella serovar Typhimurium infection than wild-type mice. These data suggest that the AI-3/Epi/NE signaling system is a key factor during Salmonella serovar Typhimurium pathogenesis in vitro and in vivo. Elucidation of the role of this interkingdom signaling system in Salmonella serovar Typhimurium should contribute to a better understanding of the complex interplay between the pathogen and the host during infection.

Resistance mechanism development to the topoisomerase-I inhibitor Hoechst 33342 byLeishmania donovani

Parasitology ◽  
2005 ◽  
Vol 131 (2) ◽  
pp. 197-206 ◽  
Author(s):  
J.-F. MARQUIS ◽  
I. HARDY ◽  
M. OLIVIER
Keyword(s):  
Mammalian Cells ◽  
Topoisomerase I ◽  
Resistance Mechanism ◽  
Resistance Mechanisms ◽  
Drug Accumulation ◽  
Wild Type ◽  
Hoechst 33342 ◽  
In The Wild ◽  
Energy Dependent

The bisbenzimidazole compound Hoechst 33342 (Ho342) has been identified as a specific Topoisomerase-I (Topo-I) inhibitor in mammalian cells. More recently, we have reported the ability of Ho342 to targetL. donovaniTopo-I, leading to parasite growth inhibitionin vitroby mechanisms involving DNA breakage and apoptosis-like phenomenon. As the Ho342 lead molecule (2,5′-Bi-1H-benzimidazole) can be used as a starting structure for derivative compounds more effective againstLeishmania, defining the Ho342 resistance mechanism(s) inLeishmaniarepresents an important strategic tool. In the present study, we selected resistant parasites to Ho342 (LdRHo.300). While we observed an increase of the Topo-I gene expression correlated by a higher Topo-I DNA relaxation activity, the Topo-I genes (LdTOP1AandLdTOP1B) sequencing did not reveal any mutation for the resistant parasites. Moreover, our results on Ho342 cellular accumulation suggested the presence of a potential energy-dependent Ho342 transporter in the wild-type parasite, and that an alteration of this transporter has occurred inLdRHo.300, leading to an altered drug accumulation. Collectively, Ho342 resistance characterization provided results supporting that the resistance developed byLdRHo.300involves complex mechanisms, most likely dominated by an altered drug accumulation, providing new insight in the Ho342 resistance mechanisms.

scholarly journals Evaluation of the in vitro Activity and in vivo Efficacy of Anidulafungin-Loaded Human Serum Albumin Nanoparticles Against Candida albicans

2021 ◽  
Vol 12 ◽  
Author(s):  
Yu Zhang ◽  
Yan-Chao Liu ◽  
Si-Min Chen ◽  
Hui Zong ◽  
Wei-Tong Hou ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Water Solubility ◽  
Fungal Infections ◽  
Antifungal Susceptibility ◽  
Fungal Species ◽  
Invasive Infection

Recent decades have seen a significant increase in invasive fungal infections, resulting in unacceptably high mortality rates. Anidulafungin (AN) is the newest echinocandin and appears to have several advantages over existing antifungals. However, its poor water solubility and burdensome route of administration (i.e., repeated, long-term intravenous infusions) have limited its practical use. The objective of this study was to develop anidulafungin-loaded Human Serum Albumin (HSA) nanoparticles (NP) so as to increase both its solubility and antifungal efficacy. HSA was reduced using SDS and DTT, allowing liberation of free thiols to form the intermolecular disulfide network and nanoassembly. Reduced HSA was then added to MES buffer (0.1 M, pH 4.8) and magnetically stirred at 350 rpm and 25°C with AN (m/m 50:1) for 2 h to form nanoparticles (AN NP). We next performed routine antifungal susceptibility testing of Candida strains (n = 31) using Clinical and Laboratory Standards Institute (CLSI) methodologies. Finally, the in vivo efficacy of both AN and AN NP was investigated in a murine model of invasive infection by one of the most common fungal species—C. albicans. The results indicated that our carrier formulations successfully improved the water solubility of AN and encapsulated AN, with the latter having a particle size of 29 ± 1.5 nm with Polymer dispersity index (PDI) equaling 0.173 ± 0.039. In vitro AN NP testing revealed a stronger effect against Candida species (n = 31), with Minimum Inhibitory Concentration (MIC) values 4- to 32-fold lower than AN alone. In mice infected with Candida and having invasive candidiasis, we found that AN NP prolonged survival time (P < 0.005) and reduced fungal burden in kidneys compared to equivalent concentrations of free drug (P < 0.0001). In conclusion, the anidulafungin nanoparticles developed here have the potential to improve drug administration and therapeutic outcomes for individuals suffering from fungal diseases.

scholarly journals Quantitative Microplate-Based Growth Assay for Determination of Antifungal Susceptibility of Histoplasma capsulatum Yeasts

2015 ◽  
Vol 53 (10) ◽  
pp. 3286-3295 ◽  
Author(s):  
Kristie D. Goughenour ◽  
Joan-Miquel Balada-Llasat ◽  
Chad A. Rappleye
Keyword(s):  
Fungal Pathogens ◽  
Histoplasma Capsulatum ◽  
Dynamic Range ◽  
Antifungal Susceptibility ◽  
Fungal Growth ◽  
Invasive Fungal Disease ◽  
Phylogenetic Groups ◽  
Morphological Form ◽  
Content Type

Standardized methodologies for determining the antifungal susceptibility of fungal pathogens is central to the clinical management of invasive fungal disease. Yeast-form fungi can be tested using broth macrodilution and microdilution assays. Reference procedures exist forCandidaspecies andCryptococcusyeasts; however, no standardized methods have been developed for testing the antifungal susceptibility of yeast forms of the dimorphic systemic fungal pathogens. For the dimorphic fungal pathogenHistoplasma capsulatum, susceptibility to echinocandins differs for the yeast and the filamentous forms, which highlights the need to employHistoplasmayeasts, not hyphae, in antifungal susceptibility tests. To address this, we developed and optimized methodology for the 96-well microtiter plate-based measurement ofHistoplasmayeast growthin vitro. Using optical density, the assay is quantitative for fungal growth with a dynamic range greater than 30-fold. Concentration and assay reaction time parameters were also optimized for colorimetric (MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction) and fluorescent (resazurin reduction) indicators of fungal vitality. We employed this microtiter-based assay to determine the antifungal susceptibility patterns of multiple clinical isolates ofHistoplasmarepresenting different phylogenetic groups. This methodology fulfills a critical need for the ability to monitor the effectiveness of antifungals onHistoplasmayeasts, the morphological form present in mammalian hosts and, thus, the form most relevant to disease.

scholarly journals Baseline Sensitivities of Fungal Pathogens of Fruit and Foliage of Citrus to Azoxystrobin, Pyraclostrobin, and Fenbuconazole

Plant Disease ◽  
2005 ◽  
Vol 89 (11) ◽  
pp. 1186-1194 ◽  
Author(s):  
S. N. Mondal ◽  
Alka Bhatia ◽  
Turksen Shilts ◽  
L. W. Timmer
Keyword(s):  
Mycelial Growth ◽  
Fungal Pathogens ◽  
Fungal Species ◽  
Brown Spot ◽  
Colletotrichum Acutatum ◽  
Fungicide Sensitivity ◽  
Fruit Drop ◽  
Coefficients Of Variation ◽  
Foliar Pathogens

The baseline sensitivities for mycelial growth of foliar fungal pathogens of citrus, Colletotrichum acutatum, Alternaria alternata, Elsinoe fawcettii, Diaporthe citri, and Mycosphaerella citri, the causal agents of postbloom fruit drop, brown spot of tangerine, citrus scab, melanose, and greasy spot, respectively, were determined in vitro for azoxystrobin, pyraclostrobin, and fenbuconazole. The effective dose to reduce growth by 50% (ED50 values) was determined for each pathogen-fungicide combination using five isolates from different citrus areas of Florida and eight fungicide concentrations. A discriminatory dose for each combination was selected near the ED50, and the range of sensitivity of 50 to 62 isolates of each fungal species was determined. The effect of salicylhydroxamic acid (SHAM) on the sensitivity of the five fungal species to azoxystrobin and pyraclostrobin was determined. Since mycelial growth of A. alternata was insensitive to azoxystrobin, the effect of that fungicide with and without SHAM on spore germination was assessed. The ED50 values for most fungal pathogens of citrus were relatively high compared with foliar pathogens of other tree crops. Values for azoxystrobin ranged from a low of 0.06 μg/ml with E. fawcettii to a high of >100 μg/ml with A. alternata. With pyraclostrobin, the values ranged from a low of 0.019 μg/ml with D. citri to a high of 0.87 μg/ml with A. alternata. With fenbuconazole, the lowest ED50 value was 0.21 μg/ml with M. citri and the highest was 1.01 μg/ml with C. acutatum, but A. alternata and D. citri were not tested. SHAM was inhibitory to all species and reduced growth of D. citri greatly. Inclusion of SHAM in the medium did not greatly affect the sensitivity of mycelial growth of these fungi to azoxystrobin or pyraclostrobin, nor did it affect the ED50 values for conidial germination of A. alternata. The coefficients of variation for the sensitivity of 50 to 62 isolates of each species to these fungi ranged from 7.3% with the pyraclostrobin-C. acutatum combination to a high of 55.0% with the fenbuconazole- M. citri combination. Discriminatory doses have been established for these pathogen- fungicide combinations that should be useful for detecting major shifts in fungicide sensitivity.

scholarly journals Epidemiology and Molecular Characterizations of Azole Resistance in Clinical and Environmental Aspergillus fumigatus Isolates from China

2016 ◽  
Vol 60 (10) ◽  
pp. 5878-5884 ◽  
Author(s):  
Yong Chen ◽  
Zhongyi Lu ◽  
Jingjun Zhao ◽  
Ziying Zou ◽  
Yanwen Gong ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Antifungal Susceptibility ◽  
Public Health Problem ◽  
Wide Distribution ◽  
Resistance Mechanisms ◽  
Azole Resistance ◽  
Common Mutation ◽  
Content Type ◽  
Environmental Resistance ◽  
Resistant Strains

ABSTRACTAzole resistance inAspergillus fumigatushas emerged as a worldwide public health problem. We sought here to demonstrate the occurrence and characteristics of azole resistance inA. fumigatusfrom different parts of China. A total of 317 clinical and 144 environmentalA. fumigatusisolates from 12 provinces were collected and subjected to screening for azole resistance. Antifungal susceptibility,cyp51Agene sequencing, and genotyping were carried out for all suspected azole-resistant isolates and a subset of azole-susceptible isolates. As a result, 8 (2.5%) clinical and 2 (1.4%) environmentalA. fumigatusisolates were identified as azole resistant. Five azole-resistant strains exhibit the TR34/L98H mutation, whereas four carry the TR34/L98H/S297T/F495I mutation in thecyp51Agene. Genetic typing and phylogenetic analysis showed that there was a worldwide clonal expansion of the TR34/L98H isolates, while the TR34/L98H/S297T/F495I isolates from China harbored a distinct genetic background with resistant isolates from other countries. High polymorphisms existed in thecyp51Agene that produced amino acid changes among azole-susceptibleA. fumigatusisolates, with N248K being the most common mutation. These data suggest that the wide distribution of azole-resistantA. fumigatusmight be attributed to the environmental resistance mechanisms in China.

scholarly journals SUSCEPTIBILITY TEST FOR FUNGI: CLINICAL AND LABORATORIAL CORRELATIONS IN MEDICAL MYCOLOGY

2015 ◽  
Vol 57 (suppl 19) ◽  
pp. 57-64 ◽  
Author(s):  
Ana ALASTRUEY-IZQUIERDO ◽  
Marcia S.C. MELHEM ◽  
Lucas X. BONFIETTI ◽  
Juan L. RODRIGUEZ-TUDELA
Keyword(s):  
Susceptibility Testing ◽  
Fungal Infections ◽  
Antifungal Susceptibility ◽  
Fungal Species ◽  
Medical Mycology ◽  
Candida Spp ◽  
Disease Epidemiology ◽  
Resistant Strains ◽  
European Standards

SUMMARYDuring recent decades, antifungal susceptibility testing has become standardized and nowadays has the same role of the antibacterial susceptibility testing in microbiology laboratories. American and European standards have been developed, as well as equivalent commercial systems which are more appropriate for clinical laboratories. The detection of resistant strains by means of these systems has allowed the study and understanding of the molecular basis and the mechanisms of resistance of fungal species to antifungal agents. In addition, many studies on the correlation of in vitro results with the outcome of patients have been performed, reaching the conclusion that infections caused by resistant strains have worse outcome than those caused by susceptible fungal isolates. These studies have allowed the development of interpretative breakpoints for Candida spp. and Aspergillus spp., the most frequent agents of fungal infections in the world. In summary, antifungal susceptibility tests have become essential tools to guide the treatment of fungal diseases, to know the local and global disease epidemiology, and to identify resistance to antifungals.

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