Fluconazole-resistant Candida Species Research Articles

Screening of C. auris among Candida isolates from various tertiary care institutions in Lahore by VITEK 2 and real time PCR based molecular technique.

Candida auris is a multidrug-resistant pathogen, that is a well-known cause of nosocomial infections. This pathogen is being identified using advanced diagnostic approaches and epidemiological typing procedures. In underdeveloped nations, several researchers developed and validated a low-cost approach for reliably identifying Candida auris. The goal of this study was to assess the burden of Candida auris in different teaching hospitals of Lahore and to limit its spread to minimize hospital-related illnesses. Candida isolates were obtained from various tertiary care institutions in Lahore in the form of culture on various culture plates. Sabouraud agar culture plates were used to culture the Candida spp. Fluconazole-resistant Candida species were chosen for further identification using VITEK 2 Compact ID and molecular identification using species-specific PCR assay. The current study obtained 636 Candida samples from several tertiary care institutions in Lahore. Fluconazole resistance was found in 248 (38.9%) of 636 Candida samples. No isolate was identified as Candida auris by VITEK 2 Compact ID and real-time PCR-based molecular identification. Thus with limited resources, these two methods may serve as useful screens for Candida auris. However, it should be screened all over the country to limit its spread to break the chain of nosocomial infections.

Antifungal activity of Co(II) and Cu(II) complexes containing 1,3-bis(benzotriazol-1-yl)-propan-2-ol on the growth and virulence traits of fluconazole-resistant Candida species: synthesis, DFT calculations, and biological activity

Relevant virulence traits in Candida spp. are associated with dimorphic change and biofilm formation, which became an important target to reduce antifungal resistance. In this work, Co(II) complexes containing a benzotriazole derivative ligand showed a promising capacity of reducing these virulence traits. These complexes exhibited higher antifungal activities than the free ligands against all the Candida albicans and non-albicans strains tested, where compounds 2 and 4 showed minimum inhibitory concentration values between 15.62 and 125 μg mL−1. Moreover, four complexes (2–5) of Co(II) and Cu(II) with benzotriazole ligand were synthesized. These compounds were obtained as air-stable solids and characterized by melting point, thermogravimetric analysis, infrared, Raman and ultraviolet/visible spectroscopy. The analysis of the characterization data allowed us to identify that all the complexes had 1:1 (M:L) stoichiometries. Additionally, Density Functional Theory calculations were carried out for 2 and 3 to propose a probable geometry of both compounds. The conformer Da of 2 was the most stable conformer according to the Energy Decomposition Analysis; while the conformers of 3 have a fluxional behavior in this analysis that did not allow us to determine the most probable conformer. These results provide an important platform for the design of new compounds with antifungal activities and the capacity to attack other target of relevance to reduce antimicrobial resistance.

Invasive Aspergillosis and Candidiasis in a Patient with Supraglottic Carcinoma undergoing Chemoradiotherapy: A Case Report

The incidence of Invasive Aspergillosis (IA) is increasing. Early diagnosis and treatment are very important to improve patient survival, especially in immunocompromised patients. The diagnosis of IA is challenging as clinical indicators are limited, and early microbiological confirmation of the infection is infrequent. Here, the authors present the case of a 43-year-old male who was undergoing radiotherapy along with concurrent chemotherapy for carcinoma supraglottis. He developed Invasive Pulmonary Aspergillosis (IPA) along with candidiasis within two weeks of therapy. Voriconazole remains the recommended therapy for patients with IA, which also acts against fluconazole-resistant Candida species.

Broad-spectrum antifungal activity of Phyllanthus niruri leaves tested against Candida species

Introduction: The emergence of resistant pathogenic microorganisms against conventional antimicrobials has become a global concern. To combat new and re-emerging infectious illnesses, new antimicrobial agents with different chemical structures and novel modes of action are required. Therefore, this study evaluated ethanolic extracts of Phyllanthus niruri for their antimicrobial activities against Candida species isolated from different clinical samples. Materials and Methods: This involved the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for the identification of Candida species. The ethanolic extraction of P. niruri leaves was examined. The agar well diffusion method was used to determine the antifungal activity of the leaf extracts against standard American Type Culture Collection strains as well as clinical isolates of Candida species. Results: Using PCR-RFLP, Candida tropicalis was found to be the most prevalent species of Candida, followed by Candida albicans, Candida glabrata, Candida krusei and Candida parapsilosis. The ethanol extract of P. nirui leaves showed good activity against all the clinical and standard strains of Candida which were comparable to the standard drug fluconazole. The activity of P. niruri against C. albicans was highest followed by C. parapsilosis, C. tropicalis, C. glabrata and C. krusei at a concentration of 100 mg/ml. Conclusion: The findings of this study support the use of P. niruri plant to treat Candida infections, particularly fluconazole-resistant Candida species.

Detection of ERG11 gene in fluconazole resistant urinary candida isolates

Candida species resistant to fluconazole and voriconazole were screened for the presence of ERG11gene by polymerase chain reaction (PCR). Also, the association of this gene with the demonstration of Candida virulence factors; biofilm formation, phospholipase and proteinases activities were studied. A total of 61 Candida isolates were collected from urine specimens. Candida species were identified by API 20 C Aux test. Extracellular phospholipase, secretory aspartyl proteinase and biofilm formation were determined. ERG11 gene was detected by PCR. C. albicans was identified in 34.5%, C. glabrata in 29.5% and C. tropicalis and C. krusei in 18% each. Candida species was resistant to fluconazole and voriconazole in 55.7% and 27.9%, respectively. Seventeen (50%) of fluconazole resistant Candida isolates were sensitive to voriconazole. The most frequently Candida species revealed fluconazole resistance were C. glabrata (47.1%), C. krusei (29.4%), and C. tropicalis and C. albicans (11.8% each). Biofilm formation, phospholipase and proteinase activity were determined in 41.2%, 67.6% and 35.3% of fluconazole resistant Candida isolates, respectively. Erg 11 gene was determined in 82.4% of fluconazole resistant Candida isolates and prominent in C. glabrata (93.75%), followed by C. krusei (90%), C. tropicalis (75%) and C. albicans (25%). Erg 11 gene was detected in 64.7% (11/17) of fluconazole resistant-voriconazole sensitive Candida isolates. Regarding, correlation of Erg11 gene positivity and virulence factors among fluconazole resistant Candida isolates, 34.5% exhibited biofilm formation and 62.1% and 31% showed phospholipase and proteinase activities, respectively. There were statistically significant difference concerning the association of proteinase activities and Erg 11 gene expression among fluconazole resistance Candida isolates (P=0.04). The study emphasizes the high prevalence of Erg11 gene among fluconazole resistant Candida species. There was association between the proteinase activity, fluconazole resistance and the presence of Erg11 among Candida species. Voriconazole maintains better activity towards Candida species and represent an alternative therapy.

Isolation Frequency of Fluconazole-Resistant Candida Species From Cockroaches: A Cross-Sectional Study From a National Hospital in Dar es Salaam, Tanzania.

Background: Cockroaches are common pests in homes and hospitals. They cause allergic reactions in some individuals and are potential vectors for various infectious pathogens. The study investigated the extent to which hospital cockroaches act as vectors and reservoirs of medically important fungal pathogens on their external surfaces.Methods: Cockroaches were captured from the selected hospital locations including the burn unit, adult surgical wards, pediatric oncology wards, intern hostel kitchen, and the central kitchen of a national referral teaching hospital in Tanzania. Normal saline washings from the external surface of cockroaches were cultured on standard mycological media to facilitate isolation and identification of medically important molds and yeasts. The susceptibility of Candida species isolates to fluconazole was tested using the Clinical and Laboratory Standards Institute (CLSI) M27-A3 microdilution method.Results: A total of 69 cockroaches were captured from various hospital sites between February and April 2017. All cockroaches captured were shown to carry medically important fungi. A total of 956 medically important fungi were isolated; 554 (57.9%) were of Candida species, 222 (23.2%) were of Aspergillus species, 30 (3.1%) were of​​​​​​​ Cladosporium species, 17 (1.8%) were of​​​​​​​ Rhizopus species, 11 (1.2%) were of​​​​​​​ Geotrichum species, nine (0.9%) were of​​​​​​​ Penicillium species, seven (0.7%) were of​​​​​​​ Alternaria species, six (0.6%) were of​​​​​​​ Fusarium species, three (0.3%) were of​​​​​​​ Mucor species, and 97 (10.1%) were of other species. Of the Aspergillus species, Aspergillus fumigatus (111, 50.0%) was the most commonly isolated, followed by Aspergillus niger (35, 15.8%) among the Aspergillus isolates. Out of the 103 selected isolates, 18 (17.5%) of the Candida isolates normally not intrinsically resistant to fluconazole demonstrated resistance to this drug. Resistance was most frequently found in Candida parapsilosis (3, 30%), Candida pseudotropicalis (10, 23.8%), and Candida glabrata (2, 18.2%). The isolates with the least proportion of resistance to fluconazole were Candida albicans (2, 6.3%).Conclusion: Cockroaches from this hospital may act as reservoirs of medically important opportunistic fungi exhibiting resistance to fluconazole.

Effect of copper on the antifungal activity of disulfiram (Antabuse®) in fluconazole-resistant Candida strains.

The effect of copper on the antifungal activity of disulfiram was evaluated against fluconazole-resistant Candida species. The study establishes that copper addition confers greater inhibition of disulfiram-treated Candida cultures, but the combination antagonizes the killing effects of disulfiram.

Effectiveness of Fluconazole Prophylaxis in a Targeted High-Risk Group in a Surgical Intensive Care Unit.

Background: Candidemia is an important nosocomial infection in intensive care units (ICUs), with total parenteral nutrition (TPN) a well-recognized risk factor. Antifungal prophylaxis may be an effective intervention to prevent candidemia in high-risk patients. In this report, the effectiveness of fluconazole prophylaxis was examined in patients located in a combined surgical-neurosurgical ICU serving an urban Level 1 trauma center who were receiving prolonged courses of TPN. Methods: Fluconazole was administered prophylactically for patients receiving TPN for more than six days. Rates of candidemia during the intervention were compared with those prior to the intervention. Results: During the 27-month pre-intervention period, seven episodes of candidemia occurred during 1,277 days of parenteral nutrition therapy. During the 17-month post-intervention period, there were zero episodes during 852 days of therapy (p = 0.03). Similarly, during the pre-intervention period, there were six episodes of candidemia during 867 high-risk days of therapy, compared with zero during 643 days of high-risk therapy in the post-intervention period (p = 0.04). The rates of bacteremia did not change, and emergence of fluconazole-resistant Candida species was not evident. Conclusions: At our surgical ICU, this fluconazole prophylaxis was associated with a significant decrease in the number of patients with candidemia, without emergence of resistant species.

Antifungal Resistance Pattern of Candida Species Isolated from High Vaginal Swabs of Women Attending a Hospital in Enugu State, Nigeria

There is an increase in non-albicans Candida (NAC) vulvovaginal candidiasis which is attributed to overuse of antifungal therapy and this has led to antifungal resistance. This study was aimed at determining the antifungal resistance pattern of some clinical isolates of Candida species. Eighty-eight (88) isolates were used which included Candida tropicalis (34), Candida Parapsilosis (21), Candida albicans (20), Candida krusei (7) and Candida glabrata (6). The drugs used were Fluconazole (25µg), Ketoconazole (10µg), Voriconazole (1µg), Nystatin (100Units), Amphotericin B (20µg), Flucytosine (1µg), Clotrimazole (10µg) and Itraconazole (50µg). The susceptibility testing was carried out using the M44-A standard method for yeast disk diffusion testing. Results showed that the percentages of Candida species resistant to Fluconazole, Ketoconazole, Voriconazole, Amphotericin B, Flucytosine, Clotrimazole and Itraconazole and Nystatin were 52.3%, 61.9%, 35.2%, 19.3%, 86.4%, 34.1%, 45.5% and 44.3%, with inhibition zone diameters ≤14mm, ≤20mm, ≤13mm, <10mm, ≤11mm, ≤11mm, ≤13mm and no inhibition zone diameter respectively. Candida krusei was the most resistant species with 100% resistance to each of Fluconazole, Ketoconazole and Flucytosine. Candida tropicalis was the species with the highest susceptibility (79.4%) to Amphotericin B followed by Candida parapsilosis with inhibition zone diameters ≥15mm. While Candida glabrata showed 100% resistance to each of Flucytosine and Itraconazole, Candida albicans showed 100% resistance to Flucytosine only. Candida glabrata was the only Candida species with 0% resistance to Amphotericin B. The drug to which most of the Candida species were susceptible was Amphotericin B followed by Voriconazole while Flucytosine was the drug with the highest resistance followed by Ketoconazole and Fluconazole. The highest number of susceptible-dose dependent Candida isolates was observed with Ketoconazole (25%), followed by Clotrimazole and Itraconazole, each recording 23.9%. Based on the findings of the present study, Voriconazole is recommended for vaginal candidiasis especially in the study area and also especially for infections caused by Fluconazole-resistant Candida species. This suggests that routine sensitivity testing is pertinent to guiding the choice of antifungal therapy. Thus, indiscriminate use of antifungal drugs should be avoided to reduce the development and spread of resistance.

Screening the antifungal activities of monoterpenes and their isomers against Candida species.

Increasing the occurrence of non-albicans Candida species with intrinsic or acquired resistance to antifungals as well as the emergence of multidrug Candida species coupled with the limited antifungal agents challenges the treatment of candidiasis. Consequently, a class of secondary metabolites of plants exhibiting decent antifungal activity. Therefore, this study aimed to evaluate the antifungal potential of various monoterpenes including Carvone, Limonene, Pinene, Menthone, Menthol, Camphor, Thujone, Citronellol, and Piperitone against standard and clinical isolates of Candida. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of compounds were determined, using the broth Microdilution method based on M27-A3 protocol documented by clinical laboratory standard institute (CLSI). Amongst the tested monoterpenes, oxygenated terpenoids showed strong antifungal activity. Specifically, alcoholic terpenoids such as (±)-Citronellol possess more efficacy than the corresponding ketonic ones with MICs ranging from 0·03 to 2·00μlml-1 (0·16-10·80mmoll-1 ). Among the examined yeasts, Candida tropicalis was the most susceptible species to (±)-Citronellol. Moreover, the examined monoterpenes successfully inhibited the growth of fluconazole-resistant Candida species. Moreover, statistical analysis showed no statistically significant difference between the (+) and (-) isomers, except for (±)-α-Pienene and (±) Menthone (ρ-value<0·05). Among the tested monoterpenes, (±)-Citronellol was the most potent compounds followed by (+)-α-Pinene and Menthol. Considering the significant antifungal activity of the examined monoterpenes, they could be used in controlling or treating candidiasis. Those potent antifungal monoterpenes with GRAS status in addition to their pleasant taste and odour make them appropriate additive or preservative compounds in food and cosmetics products. Furthermore, these data might help researchers to predict EOs antifungal activities, after determining its constituents. This study provides new information about the antifungal activities of monoterpenes and their isomers presented widely in essential oils. Screening results against pathogenic yeasts confirm the correlation between the chemical structure of tested monoterpenes and their antifungal effects. The present findings might be helpful to anticipate the antifungal activity of essential oils.

Review of the Current Management of Urinary Tract Infections due to Fluconazole-Resistant and Non-Albicans Candida Species

The management of fungal infections is challenging, especially given the continued increase in fluconazole resistance and prevalence of non-albicans Candida spp. Urinary tract infections due to non-albicans and fluconazole-resistant Candida are particularly problematic given the uncertainty in determining true infection from colonization and the limited number of antifungal agents with adequate penetration to the site of infection. While our understanding of mechanisms of resistance and pharmacokinetics and pharmacodynamics continues to improve, it is combatted by the emergence of novel, difficult-to-manage pathogens such as C. auris. The purpose of this review is to summarize recent data regarding the management of urinary tract infections due to non-albicans and fluconazole-resistant Candida spp. with a focus on mechanisms of resistance, pharmacokinetics and pharmacodynamics, and treatment strategies. Advances in molecular techniques have improved our understanding of the mechanisms responsible for antifungal resistance, especially to the echinocandins and azole antifungal classes. Additionally, increased knowledge of the pharmacokinetic and pharmacodynamic properties that govern available antifungal agents has enhanced the ability to optimize dosing, especially for extravascular fungal infections. Unfortunately, these developments in the pre-clinical arena have not been matched by additional high-quality clinical data, and therefore, the optimal management strategies for fungal UTIs remain elusive. The lack of adequate new clinical data to guide optimal management of UTIs due to fluconazole-resistant and non-albicans Candida, combined with the dearth of novel antifungal agents, leaves fluconazole, amphotericin B deoxycholate, and flucytosine as the drugs of choice for these infections. Further data regarding urinary concentrations and optimal urinary PK/PD parameters may allow for the use of other agents, such as the echinocandins, in certain situations. Several novel antifungal agents are currently in clinical development with adequate in vitro activity against fluconazole-resistant and non-albicans Candida, although their utility in the treatment of UTIs requires further investigation.

Management of Antimicrobial Agents in Abdominal Organ Transplant Patients in Intensive Care Unit.

Purpose of ReviewEarly diagnosis of infections and immediate initiation of appropriate antimicrobials are crucial in the management of patients before and after organ transplantation. We reviewed the most recent literature and guidelines in this field and organized the current recommendations for healthcare professionals caring for critically ill organ transplant recipients.Recent FindingsThe incidence of multidrug-resistant organisms is increasing. Multidrug-resistant Gram-negative bacteria comprise about 14% of organisms. Vancomycin-resistant enterococci bloodstream infections are also on the rise, as 20.5% of nosocomial enterococci are now vancomycin-resistant, changing empiric antibiotic selection. Fluconazole-resistant Candida species comprise up to 46% of cases of candidemia in hospitalized patients. Consequently, new guidelines recommend primary use of echinocandins in patients with candidemia who have moderate-to-severe disease. Finally, the incidence of emergence of ganciclovir-resistant cytomegalovirus infection in patients is 5–12%, requiring early recognition and change to alternative regimens in the case of poor response to therapy.SummaryBloodstream infections are a major cause of mortality and morbidity in solid organ transplantation. Mortality as high as 24% and 50% have been reported with sepsis and septic shock respectively. As such, bloodstream infections should be diagnosed rapidly and intravenous antibiotics should be started immediately. Appropriate resuscitation should be initiated and the number and/or dose of immunosuppressive drugs should be reduced. Proper source control must also be achieved with radiologic drainage or surgical intervention as appropriate. Initial antibiotic treatment of these patients should cover both Gram-positive organisms, especially in the presence of intravascular catheters, and Gram-negative bacteria. Echinocandins like caspofungin should also be considered especially in critically ill patients, particularly if a patient has been on total parenteral nutrition or broad-spectrum antibiotics.

Indifferent effect of nonsteroidal anti-inflammatory drugs (NSAIDs) combined with fluconazole against multidrug-resistant Candida auris.

Background and Purpose:Emergence and development of antifungal drug resistance in Candida species constitute a serious concern. Candida auris as an emerging multidrug-resistant fungus is the most important public health threat with high levels of mortality and morbidity. Almost all C. auris isolates are resistant to fluconazole, and there have been reports of elevated minimum inhibitory concentrations (MICs) to amphotericin B and echinocandins. To overcome the growing challenge of antifungal resistance, a valuable alternative option would be the use of drug combination.Materials and Methods:The present study evaluated the in vitro combination of nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen, diclofenac and aspirin with fluconazole against fluconazole-resistant C. auris in comparison to other fluconazole-resistant Candida species, including C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei originating from patients with candidiasis.Results:The MIC ranges of fluconazole-ibuprofen and fluconazole-diclofenac decreased from 32-256 to 32-128 and 16-256 µg/ml, respectively and remained the same for fluconazole-aspirin against C. auris. However, the combination of fluconazole with ibuprofen resulted in a synergistic effect for 5 strains, including C. albicans (n=2), C. tropicalis (n=1), C. glabrata (n=1), and C. krusei (n=1), by decreasing the MIC of fluconazole by 2-3 log2 dilutions.Conclusion:Although the interaction of NSAIDs with fluconazole was not synergistic against fluconazole-resistant C. auris isolates, no antagonism was observed for any combinations. Therefore, combination with newer azole agents needs to be conducted.

Invasive fungal infection in liver transplant recipients in a prophylactic era: A multicenter retrospective cohort study in Korea.

The epidemiology of invasive fungal infections (IFIs) after liver transplantation (LT) is continuing to evolve in the current era of antifungal prophylactic therapy. This multicenter retrospective cohort study aimed to evaluate the epidemiology, risk factors, and outcomes of IFIs among LT recipients in the current era.We analyzed a total of 482 LT recipients aged 18 years and older who were admitted to 3 tertiary hospitals in Korea between January 2009 and February 2012.Twenty-four episodes of IFIs occurred in 23 patients (4.77%; 23/482). Of these episodes, 20 were proven cases and 4 were probable cases according to EORTC/MSG criteria. Among these cases, IFI developed within 30 days of transplantation in 47.8% of recipients, from 31 to 180 days in 34.8% of recipients, and from 181 to 365 days in 17.4% of recipients. The most common isolates were Candida species (n = 12, 52.2%; Candida albicans, 6 cases; Candida tropicalis, 1 case; Candida glabrata, 1 case; Candida parapsilosis, 1 case; and unspecified Candida species, 1 case) and Aspergillus species (n = 7, 30.4%). The mortality in patients with IFIs was significantly higher than that in patients without IFIs (47.83% [11/23] vs 7.18% [33/459], P < .001). The incidence of late-onset IFIs is increasing in the antifungal prophylactic era, and fluconazole-resistant non-albicans Candida species have not yet emerged in Korea.

K21 Compound, a Potent Antifungal Agent: Implications for the Treatment of Fluconazole-Resistant HIV-Associated Candida Species.

Background/Objectives: With mucocutaneous candidiasis being highly prevalent in HIV patients, the emergence of fluconazole-resistant Candida species forms a major challenge in treating and eradicating these infections. The objective of this study was to establish the antifungal activity of K21, a membrane-rupturing antimicrobial compound derived from a silica quaternary ammonium compound (SiQAC) with tetraethoxysilane (TEOS).Methods: The study sample included 81 Candida species of which 9 were type strains and 72 were clinical isolates. Minimum inhibitory concentrations, synergy, fractional inhibitory concentration index (FICI), and time kill assays were determined by broth microdilution. Electron microscopy (EM) was used to determine the qualitative changes brought about after treatment with K21.Results: K21 inhibited the growth of all fluconazole-resistant and susceptible Candida strains with only 2 h of exposure required to effectively kill 99.9% of the inoculum, and a definite synergistic effect was observed with a combination of K21 and fluconazole. EM demonstrated the presence of two forms of extracellular vesicles indicative of biofilm formation and cell lysis.Conclusion: The study established the efficacy of K21 as an antifungal agent and with fluconazole-resistant candidiasis on the increase, the development of K21 can provide a promising alternative to combat acquired drug resistance.

An update on the application of nano-scaled carriers against fluconazole-resistant Candida species: nanostructured lipid carriers or solid lipid nanoparticles?

Background and Purpose:Encapsulation can lead to improved efficacy and safety of antifungal compounds. The attention of scientists has recently turned to biocompatible lipids as the carriers for the delivery of antifungal drugs, such as fluconazole. Although several research reports have already been published on fluconazole loaded solid lipid nanoparticles (FLZ-SLNs) and fluconazole loaded nanostructured lipid carriers (FLZ-NLCs), the possible advantages of NLCs over SLNs have not yet been fully established. Studies performed so far have given several contradictory results.Materials and Methods:Both formulations of fluconazole were synthesized using probe ultrasonication method and the characteristics were analyzed. Antifungal susceptibility testing (AFST) was performed with FLZ, FLZ-SLNs, and FLZ-NLCs using CLSI document M60 against some common fluconazole-resistant Candida species.Results:A significant decrease was observed in minimum inhibitory concentration values when both formulations were applied. Nonetheless, FLZ-NLCs were significantly more effective (P<0.05). However, three species groups were not statistically different in terms of the activity of FLZ-NLCs.Conclusion:Based on the obtained results, FLZ-NLCs could reverse the azole-resistance phenomenon in the most common Candida species more effectively, as compared to FLZ-SLNs.

Original Research Article (Experimental): Targeting fungal menace through copper nanoparticles and Tamrajal

BackgroundWHO reports, an escalation of antibiotic resistance in opportunistic pathogens like Candida. Tamrajal, i.e., water stored in copper vessels has been proclaimed as health elixir by ancient Ayurveda. Vis-a-Vis the use of copper contact surfaces and nanoparticles has gained significance for their antimicrobial effects. It thus seems imperative to examine copper nanoparticles and tamrajal as promising alternatives to existing antifungals. ObjectiveThis study not only assessed the influence of Tamrajal and copper nanoparticles on the morphological alterations of the Candida and its biofilm forming ability, but also on their ability to destroy preformed biofilms. Materials and methodsCopper oxide nanoparticles as well as Tamrajal were evaluated as complementary as well as stand-alone antimicrobial agents. ‘Time kill assay’ and ‘germ tube inhibition test’ were performed as end-point analysis for pathogenesis, while biofilm quantification, performed to assess the colonizing capability of Candida. Scanning Electron Microscope was used for visualizing the cells, whilst ICP-AES to determine the copper concentration. Results92–100% cytotoxicity to the fluconazole resistant Candida species was observed with copper oxide nanoparticles as well as tamrajal during 24hr time kill assay. The study also confirmed complete germ tube inhibition by copper in both its forms in addition to the reduction in the biofilm production. ConclusionCompared to the classes of antifungals like azoles, echinocandins etc, copper based anti-candidal agents highlight a potential way to combat resistant candidiasis. The possibility of accumulation of NP resulting in cytotoxicity puts tamrajal as the choice due to its efficacy as well as non-toxicity as per the EPA.

Fungicidal versus Fungistatic activity of five Iranian essences against fluconazole resistant Candida species

Introduction: Antifungal resistant is one of the causes of high mortality rates during invasive candidiasis. Since development of new antifungal agents is limited, researchers have focused on natural products including essential oils (EOs) with antifungal properties. In immunocompromised patients fungicidal activity is of benefit. This study was designed to evaluate chemical composition and fungicidal/fungistatic activities of five Iranian EOs and against fluconazole resistant Candida species.Methods: To determine chemical composition of EOs gas chromatography-mass spectroscopy (GC/MS) was employed. Fluconazole resistant Candida species were chosen and minimum inhibitory concentration (MIC) values of studied EOs were determined by broth microdilution method. Minimum fungicidal concentration (MFC) was determined as the lowest concentration with no fungal growth on solid media. Fungicidal activity was calculated by MFC/MIC ratio.Results: The results showed that C. albicans and C. tropialis isolates were susceptible to itraconazole (ITC) and voriconazole (VRC) while one species of C. glabrata and C. krusei each was resistant to itraconzaole; and itraconazole resistant C. glabrata isolate was resistance to voriconzaole as well. Among tested EOs, the ones from Cinnamomum cayennense, Origanum majorana var. majoranoides and Andropogon citratus had the highest anti-Candida activity. Artemisia aromatica A. Nelson had the highest MIC value against Candida isolates. All EOs in this study had fungicidal activity.Conclusion: In general, the tested natural compounds are suitable to be used as anti-Candida. However more studies are needed on each chemical compound to evaluate its antifungal activity alone or in combination with other agents.

Effect of fluconazole prophylaxis on Candida fluconazole susceptibility in premature infants.

Extremely premature infants are at high risk of developing invasive candidiasis; fluconazole prophylaxis is safe and effective for reducing invasive candidiasis in this population but further study is needed. We sought to better understand the effect of prophylactic fluconazole on a selection of fluconazole-resistant Candida species. We evaluated the susceptibility to fluconazole of Candida isolates from premature infants (<750 g birth weight) enrolled in a multicentre, randomized, placebo-controlled trial of fluconazole prophylaxis. Candida species were isolated through surveillance cultures at baseline (study day 0-7), period 1 (study day 8-28) and period 2 (study day 29-49). Fluconazole MICs were determined for all Candida isolates. Three hundred and sixty-one infants received fluconazole (n = 188) or placebo (n = 173). After the baseline period, Candida colonization was significantly lower in the fluconazole group compared with placebo during periods 1 (5% versus 27%; P < 0.001) and 2 (3% versus 27%; P < 0.001). After the baseline period, two infants (1%) were colonized with at least one fluconazole-resistant Candida in each group. Median fluconazole MIC was similar in both treatment groups at baseline and period 1. However, in period 2, median MIC was higher in the fluconazole group compared with placebo (1.00 versus 0.50 mg/L, P = 0.01). There was no emergence of resistance observed and no patients developed invasive candidiasis with a resistant Candida isolate. Fluconazole prophylaxis decreased Candida albicans and 'non-albicans' Candida colonization and was associated with a slightly higher fluconazole MIC for colonizing Candida isolates.

Antifungal Activity, Gas Chromatographic-Mass Spectrometric Analysis And in silico Study of Punica Granatum Peel Extracts Against Fluconazole Resistant Strains of Candida Species

Candida species is the common cause of opportunistic fungal infections all over the world with increased mortality and morbidity especially in immunosuppressed patients. Fluconazole is the first line therapy for candidiasis. The antifungal resistance pattern in high-risk patients is a major concern. The present study was aimed to assess the anticandidal activity of Punica granatum peel against fluconazole resistant Candida species isolated from HIV patients. Ethanol, chloroform, petroleum ether and aqueous extracts of the peel of P. granatum were evaluated against standard strains of Candida spp. and fluconazole resistant clinical isolates by agar diffusion and broth dilution techniques. The GC-MS analysis of the extracts was performed to identify the phytochemicals present in it. The predominant phytochemical was subjected to molecular docking study to determine its binding efficacy with lanosterol 14-alpha demethylase. P. granatum peel extracts showed excellent anticandidal activity with ethanol extract exhibiting the most inhibitory activity. C. albicans and C. krusei were the most inhibited and C. parapsilosis was the least inhibited species. The GC-MS analysis of the ethanol extract identified five predominant phytochemicals. On docking studies, the five phytochemicals showed a good binding to the lanosterol 14-alpha demethylase. The present study is the first report on the antifungal activity of various extracts of P. granatum against fluconazole resistant Candida isolates. Ethanol extract of P. granatum peel showed excellent anticandidal activity against fluconazole resistant Candida spp. Hence, it can be explored further to identify a potential drug candidate.