scholarly journals Assessment of the Effect of Amphotericin B on the Vitality of Candida albicans

1999 ◽  
Vol 43 (5) ◽  
pp. 1034-1041 ◽  
Author(s):  
Robert S. Liao ◽  
Robert P. Rennie ◽  
James A. Talbot
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Total Population ◽  
Antifungal Susceptibility ◽  
Membrane Integrity ◽  
Quantitative Information ◽  
Fungal Cell ◽  
Fluorescent Indicators ◽  
Qualitative And Quantitative ◽  
Time Kill

ABSTRACT The processes involved in cell death are complex, and individual techniques measure specific fractions of the total population. The interaction of Candida albicans with amphotericin B was measured with fluorescent probes with different cellular affinities. These were used to provide qualitative and quantitative information of physiological parameters which contribute to fungal cell viability. SYBR Green I and 5,(6)-carboxyfluorescein were used to assess membrane integrity, and bis-(1,3-dibutylbarbituric acid)trimethine oxonol and 3,3-dihexyloxacarbocyanine iodide were used to evaluate alterations in membrane potential. The fluorescent indicators were compared with replication competency, the conventional indicator of viability. By using these tools, the evaluation of the response of C. albicans to amphotericin B time-kill curves delineated four categories which may represent a continuum between alive and dead. The data showed that replication competency (CFU per milliliter) as determined by conventional antifungal susceptibility techniques provided only an estimate of inhibition. Interpretation of fluorescent staining characteristics indicated that C. albicans cells which were replication incompetent after exposure to greater than 0.5 μg of amphotericin B per ml still maintained degrees of physiological function.

scholarly journals Sublethal Injury and Resuscitation of Candida albicans after Amphotericin B Treatment

2003 ◽  
Vol 47 (4) ◽  
pp. 1200-1206 ◽  
Author(s):  
Robert S. Liao ◽  
Robert P. Rennie ◽  
James A. Talbot
Keyword(s):  
Cell Death ◽  
Candida Albicans ◽  
Amphotericin B ◽  
Antifungal Agents ◽  
Sybr Green ◽  
Sequential Treatment ◽  
Tetrazolium Salt ◽  
Fungal Cell ◽  
Sublethal Injury

ABSTRACT Amphotericin B treatment was previously shown to inhibit Candida albicans reproduction and reduce the fluorescence of vitality-specific dyes without causing a corresponding increase in the fluorescence of the mortality-specific dyes bis-(1,3-dibutylbarbituric acid)trimethine oxonol and SYBR Green Ι. In the present study, we have confirmed these results and have shown that the numbers of CFU are reduced by 99.9% by treatment with 0.5 μg of amphotericin B per ml for 10 h at 35°C. This reduction was not due to fungal cell death. First, the level of reduction of the tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide increased in the presence of concentrations of amphotericin B that caused greater than 90% reductions in the numbers of CFU. Second, fungal cells treated with amphotericin B at a concentration of 0.5 μg/ml were resuscitated by further incubation at 22°C for 15 h in the continued presence of amphotericin B. Third, recovery of the ability to replicate was prevented by sequential treatment with 20 μg of miconazole per ml, which also increased the fluorescence of mortality-specific dyes to near the maximal levels achieved with 0.9 μg of amphotericin B per ml. Sequential treatment with fluconazole and flucytosine did not increase the levels of staining with the mortality-specific dyes. Itraconazole was less effective than ketoconazole, which was less effective than miconazole. The practice of equating the loss of the capacity of C. albicans to form colonies with fungal cell death may give incorrect results in assays with amphotericin B, and the results of assays with caution with other antifungal agents that are lipophilic or that possess significant postantifungal effects may need to be interpreted.

scholarly journals Flow Cytometry Antifungal Susceptibility Testing of Pathogenic Yeasts other than Candida albicans and Comparison with the NCCLS Broth Microdilution Test

2000 ◽  
Vol 44 (10) ◽  
pp. 2752-2758 ◽  
Author(s):  
Rama Ramani ◽  
Vishnu Chaturvedi
Keyword(s):  
Flow Cytometry ◽  
Candida Albicans ◽  
Amphotericin B ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Antifungal Susceptibility Testing ◽  
Broth Microdilution ◽  
Nitrogen Base ◽  
Microdilution Method ◽  
Broth Microdilution Method

ABSTRACT Candida species other than Candida albicansfrequently cause nosocomial infections in immunocompromised patients. Some of these pathogens have either variable susceptibility patterns or intrinsic resistance against common azoles. The availability of a rapid and reproducible susceptibility-testing method is likely to help in the selection of an appropriate regimen for therapy. A flow cytometry (FC) method was used in the present study for susceptibility testing ofCandida glabrata, Candida guilliermondii,Candida krusei, Candida lusitaniae,Candida parapsilosis, Candida tropicalis, andCryptococcus neoformans based on accumulation of the DNA binding dye propidium iodide (PI). The results were compared with MIC results obtained for amphotericin B and fluconazole using the NCCLS broth microdilution method (M27-A). For FC, the yeast inoculum was prepared spectrophotometrically, the drugs were diluted in either RPMI 1640 or yeast nitrogen base containing 1% dextrose, and yeast samples and drug dilutions were incubated with amphotericin B and fluconazole, respectively, for 4 to 6 h. Sodium deoxycholate and PI were added at the end of incubation, and fluorescence was measured with a FACScan flow cytometer (Becton Dickinson). The lowest drug concentration that showed a 50% increase in mean channel fluorescence compared to that of the growth control was designated the MIC. All tests were repeated once. The MICs obtained by FC for all yeast isolates except C. lusitaniae were in very good agreement (within 1 dilution) of the results of the NCCLS broth microdilution method. Paired ttest values were not statistically significant (P = 0.377 for amphotericin B; P = 0.383 for fluconazole). Exceptionally, C. lusitaniae isolates showed higher MICs (2 dilutions or more) than in the corresponding NCCLS broth microdilution method for amphotericin B. Overall, FC antifungal susceptibility testing provided rapid, reproducible results that were statistically comparable to those obtained with the NCCLS method.

scholarly journals In Fungal Intracellular Pathogenesis, Form Determines Fate

mBio ◽  
2018 ◽  
Vol 9 (5) ◽  
Author(s):  
Robin C. May ◽  
Arturo Casadevall
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Membrane Integrity ◽  
Pathogenic Fungi ◽  
Morphological Transition ◽  
Physical Damage ◽  
Fungal Cell ◽  
Morphological Transitions ◽  
Pathogenic Microbes ◽  
Phagosomal Membrane

ABSTRACT For pathogenic microbes to survive ingestion by macrophages, they must subvert powerful microbicidal mechanisms within the phagolysosome. After ingestion, Candida albicans undergoes a morphological transition producing hyphae, while the surrounding phagosome exhibits a loss of phagosomal acidity. However, how these two events are related has remained enigmatic. Now Westman et al. (mBio 9:e01226-18, 2018, https://doi.org/10.1128/mBio.01226-18) report that phagosomal neutralization results from disruption of phagosomal membrane integrity by the enlarging hyphae, directly implicating the morphological transition in physical damage that promotes intracellular survival. The C. albicans intracellular strategy shows parallels with another fungal pathogen, Cryptococcus neoformans, where a morphological changed involving capsular enlargement intracellularly is associated with loss of membrane integrity and death of the host cell. These similarities among distantly related pathogenic fungi suggest that morphological transitions that are common in fungi directly affect the outcome of the fungal cell-macrophage interaction. For this class of organisms, form determines fate in the intracellular environment.

scholarly journals Candida albicans isolated from post-operative penetrating keratoplasty patient: Case report

2014 ◽  
Vol 5 (3) ◽  
pp. 116-119
Author(s):  
Abhishek Chandra ◽  
Munesh Kumar Gupta ◽  
Ragini Tilak
Keyword(s):  
Candida Albicans ◽  
Case Report ◽  
Amphotericin B ◽  
Penetrating Keratoplasty ◽  
Antifungal Susceptibility ◽  
Medical Science ◽  
Eye Drops ◽  
Gram Staining ◽  
Chlamydospore Formation ◽  
Culture Characteristics

We report a case report of Candida albicans suture infiltrate on 3rd post-op day in a 53 year female operated for penetrating keratoplasty. Candida albicans was identified by KOH mount, Gram Staining, germ tube, growth at 450C, chlamydospore formation and light green color on CHROMagar with sugar assimilation and culture characteristics. Despite being susceptible to Fluconazole by broth microdilution, patient did not respond to 0.3% fluconazole eye drops. On antifungal susceptibility testing by CLSI44A, it was susceptible to only Amphotericin B (100units). Patient was then started on 0.15% fortified amphotericin B eye drops resulting in complete resolution of infiltrates. Asian Journal of Medical Science, Volume-5(3) 2014: 116-119 http://dx.doi.org/10.3126/ajms.v5i3.8669 

scholarly journals Candida albicans isolated from denture-related stomatitis in elderly patients: Antifungal susceptibility and production of virulence attributes

2020 ◽  
Vol 1 ◽  
Author(s):  
Lourimar Viana Nascimento Franco de Sousa ◽  
Carlos Davi de Oliveira Maia ◽  
Isadora Sousa Carvalho ◽  
Juliano Meireles Prata ◽  
Larissa Carla Rodrigues Arcanjo ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Elderly Patients ◽  
Amphotericin B ◽  
Antifungal Susceptibility ◽  
Burning Sensation ◽  
Aspartic Protease ◽  
Elderly Individuals ◽  
Polystyrene Surface ◽  
Low Prevalence ◽  
Effective Drugs

AbstractDenture-related stomatitis caused by Candida spp. affects elderly individuals using partial/total prosthesis, provoking several discomforts including burning sensation and altered taste. Herein, we have studied 52 denture-wearing individuals (>60 years-old), attended at the dentistry clinic of UNIVALE, aiming to isolate Candida spp. directly from the stomatitis lesions and to evaluate their potential to produce virulence attributes. A low prevalence of denture-related stomatitis was reported in these patients (4/52; 7.7%). Candida albicans was isolated in the 4 selected patients, with the ability to form biofilm over a polystyrene surface and to produce aspartic protease, esterase and hemolysin. However, neither phospholipase nor caseinase activities were detected. Planktonic-growing yeasts were susceptible to amphotericin B and caspofungin, while the susceptibility to azoles (fluconazol, itraconazole and voriconazole) varied depending on either the isolate or antifungal. Relevantly, biofilm-forming C. albicans cells exhibited resistance to all studied antifungals. So, new effective drugs against resistant C. albicans isolates causing denture-related stomatitis are urgently required.

scholarly journals In Vitro Pharmacodynamic Properties of Three Antifungal Agents against Preformed Candida albicans Biofilms Determined by Time-Kill Studies

2002 ◽  
Vol 46 (11) ◽  
pp. 3634-3636 ◽  
Author(s):  
Gordon Ramage ◽  
Kacy VandeWalle ◽  
Stefano P. Bachmann ◽  
Brian L. Wickes ◽  
José L. López-Ribot
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Antifungal Agents ◽  
Pharmacokinetic Properties ◽  
Time Kill ◽  
Candida Albicans Biofilms

ABSTRACT We have examined the in vitro activities of fluconazole, amphotericin B, and caspofungin against Candida albicans biofilms by time-kill methodology. Fluconazole was ineffective against biofilms. Killing of biofilm cells was suboptimal at therapeutic concentrations of amphotericin B. Caspofungin displayed the most effective pharmacokinetic properties, with ≥99% killing at physiological concentrations.

scholarly journals Antifungal susceptibility of Candida albicans isolated from oral mucosa of patients with cancer

1999 ◽  
Vol 13 (3) ◽  
pp. 219-223 ◽  
Author(s):  
Lino João da COSTA ◽  
Esther Goldenberg BIRMAN ◽  
Sidney Hartz ALVES ◽  
Arlete Emily CURY
Keyword(s):  
Candida Albicans ◽  
Cancer Patients ◽  
Amphotericin B ◽  
Oral Mucosa ◽  
Antifungal Susceptibility ◽  
New Drugs ◽  
Local Conditions ◽  
Opportunistic Fungi ◽  
Sabouraud Agar

The increasing number of oral infections due to opportunistic fungi in immunocompromised patients, needs a new evaluation of the drugs in use. The susceptibility in vitro of Candida albicans strains from the oral mucosa of cancer patients to amphotericin B, ketoconazole, miconazole, fluconazole and itraconazole were evaluated. A dilution technique in YNB agar or subculture on Sabouraud agar was utilised for MIC or MFC determinations. With this methodology, the best fungicidal drug for C.albicans collected from the oral mucosa of 40 cancer patients, divided in two groups: one treated by radiotherapy and the other not, the best results were obtained with amphotericin B, presenting low values of MIC compared to azoles and MFC values. However it is important to take into consideration the utilisation in vivo of this polyenic antibiotic and the possible toxic levels necessary to achieve good results. The coexistence of other fungi and the local conditions must also be pondered with these patients, who are mostly undergoing radiotherapy. In order to achieve better results without undesirable consequences, higher levels of MIC are expected with the daily clinical use of new drugs.

scholarly journals Organism-Dependent Fungicidal Activities of Azoles

1998 ◽  
Vol 42 (11) ◽  
pp. 3018-3021 ◽  
Author(s):  
Elias K. Manavathu ◽  
Jessica L. Cutright ◽  
Pranatharthi H. Chandrasekar
Keyword(s):  
Candida Albicans ◽  
Aspergillus Fumigatus ◽  
Amphotericin B ◽  
The Other ◽  
Cellular Death ◽  
Antifungal Activities ◽  
Other Hand ◽  
Cytocidal Effect ◽  
Time Kill ◽  
Drug Free

ABSTRACT We investigated the antifungal activities of itraconazole and voriconazole on Aspergillus species by time kill studies, and the results were compared with those obtained forCandida species. Exposure of Aspergillus fumigatus conidia to varying concentrations (1.25 to 10 μg/ml) of itraconazole and voriconazole resulted in cellular death; the cytocidal effect was time and concentration dependent. In contrast, no killing of Candida albicans occurred in the presence of itraconazole and voriconazole at concentrations as high as 10 μg/ml, although candidal growth was inhibited compared to the drug-free control. Amphotericin B (1.25 to 10 μg/ml), on the other hand, killed both A. fumigatus and C. albicans. Similar results were obtained for non-A. fumigatus aspergilli and non-C. albicans Candida species. These observations indicate that both itraconazole and voriconazole are cytocidal agents for Aspergillus species but not for Candidaspecies, suggesting that azoles possess organism-dependent fungicidal activities.

scholarly journals Antifungal and Antibiofilm Activities and the Mechanism of Action of Repeating Lysine-Tryptophan Peptides against Candida albicans

2020 ◽  
Vol 8 (5) ◽  
pp. 758
Author(s):  
Gopal Ramamourthy ◽  
Jonggwan Park ◽  
Changho Seo ◽  
Hans J. Vogel ◽  
Yoonkyung Park
Keyword(s):  
Candida Albicans ◽  
Antifungal Activity ◽  
Laser Scanning ◽  
Membrane Integrity ◽  
Laser Scanning Confocal Microscopy ◽  
Health Concern ◽  
Cellular Functions ◽  
Fungal Cell ◽  
Scanning Confocal Microscopy ◽  
Keratinocyte Cell

The rapid increase in the emergence of antifungal-resistant Candida albicans strains is becoming a serious health concern. Because antimicrobial peptides (AMPs) may provide a potential alternative to conventional antifungal agents, we have synthesized a series of peptides with a varying number of lysine and tryptophan repeats (KWn-NH2). The antifungal activity of these peptides increased with peptide length, but only the longest KW5 peptide displayed cytotoxicity towards a human keratinocyte cell line. The KW4 and KW5 peptides exhibited strong antifungal activity against C. albicans, even under conditions of high-salt and acidic pH, or the addition of fungal cell wall components. Moreover, KW4 inhibited biofilm formation by a fluconazole-resistant C. albicans strain. Circular dichroism and fluorescence spectroscopy indicated that fungal liposomes could interact with the longer peptides but that they did not release the fluorescent dye calcein. Subsequently, fluorescence assays with different dyes revealed that KW4 did not disrupt the membrane integrity of intact fungal cells. Scanning electron microscopy showed no changes in fungal morphology, while laser-scanning confocal microscopy indicated that KW4 can localize into the cytosol of C. albicans. Gel retardation assays revealed that KW4 can bind to fungal RNA as a potential intracellular target. Taken together, our data indicate that KW4 can inhibit cellular functions by binding to RNA and DNA after it has been translocated into the cell, resulting in the eradication of C. albicans.

scholarly journals Antifungal pharmacodynamic characteristics of fluconazole and amphotericin B tested against Candida albicans.

1997 ◽  
Vol 41 (6) ◽  
pp. 1392-1395 ◽  
Author(s):  
M E Klepser ◽  
E J Wolfe ◽  
R N Jones ◽  
C H Nightingale ◽  
M A Pfaller
Keyword(s):  
Candida Albicans ◽  
Amphotericin B ◽  
Fungicidal Activity ◽  
Narrow Range ◽  
Growth Effects ◽  
Fungistatic Activity ◽  
Time Kill

Time-kill curves were determined for three isolates of Candida albicans tested against fluconazole and amphotericin B at multiples of the MIC. Fluconazole produced fungistatic activity, with concentration-related growth effects observed over a narrow range of concentrations. Amphotericin B exhibited fungicidal activity, with enhancement of activity over a broader range of concentrations.

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