Tropicalis Strains Research Articles

Seasonality influence on the chemical composition and antifungal activity of Psidium myrtoides O. Berg

The present study aimed to investigate the influence of seasonality from different collection periods on the chemical composition and antifungal activity of the Psidium myrtoides O. Berg essential oil (OEFPm). The OEFPm chemical composition was evaluated through gas chromatography coupled to mass spectrometry (GC/MS). The oil's antifungal activity against species from the Candida genus, in isolation and in combination with Fluconazole at a subinhibitory concentration (MFC/16), was evaluated using the broth microdilution method, while its effect on Candida albicans pleomorphism was evaluated by microculture. The results showed the compound 1–8 cineol was the major component in all the samples. The OEFPm showed significant MFC ranging from 1,024 to ≥ 16,384 μg/mL for the C. albicans, C. krusei and C. tropicalis strains with IC50 values ranging from 103.3 to 3564.5 μg/mL. The essential oils affected morphological transition at all tested concentrations in the C. albicans morphological alteration assays. The results from this study show the collection period affected the chemical composition and the antimicrobial activity of the essential oil. A significant variation was documented for most chemical components and the biological activity of the seasonally collected samples, in addition to variations in morphological transition inhibition, an important virulence factor for the genus.

Ocena oddziaływania na grzyby drożdżopodobne olejku szałwiowego (Oleum Salviae)

Introduction. The essential oils were known and used in ancient Times. The oil and extracts of sage are utilized in folk medicine and for food condiment, in cosmetics and perfumes. Sage (Salvia officinalis) belonging to the Labiatae family. Plant grow all over the world. Sage has a number of properties in it antiflogistic and antimicrobial. The oil is used for the treatment of different kind of diseases, including bronchitis, cough, arthritis, rheumatism, ulcers, inflammation of skin, alimentary tract and in Alzheimer’s disease. Etheric oil contain following compounds: α- and β-thujone, 1,8-cineole, camphor, borneol, α-pinene, β-pinene, β-caryophyllene, β-sabinene, limonene, α-humulene, myrcene, α-terpineol, viridiflorol and camphene. The chemical compounds of the oil have antioxidant and antimicrobial properties. Aim. The aim of the dates was to evaluate the susceptibility of yeastlike fungi to sage. Material and methods. A total 30 strains of yeastlike fungi isolated from patients with oral candidosis was tested. The strains were identified with system API 20 C AUX (BioMèrieux), production chlamydospore and pseudohyphe. The susceptibility (MIC) yeastlike fungi to sage oil was determined by means plate dilution technique in Sabouraud’s agar. The suspension contained 105 CFU per spot were spread Steers replicator over the surface of agar containing oil or without sage agar plates (strains growth control). Inoculated agar plates were incubated in aerobic conditions at 37°C for 24-48 hrs. The MIC was defined as a lowest concentration of the oil inhibited growth of fungal strains. Results. The results indicated that the sage oil was active against yeastlike fungi in concentrations 0.5-≥ 2.0 mg/ml. The MIC for 66% strains from genus Candida albicans ≥ 2.0 mg/ml. Similarly C. krusei strains were susceptible in range 0.5-≥ 2.0 mg/ml. The oil was less active towards C. glabrata and C. tropicalis strains (MIC 1.0-≥ 2.0 mg/ml). The most susceptible were the strains from genus of C. parapsilosis. The growth was inhibited within the range from 0.5 to 1.0 mg/ml. The strain from genus of Rhodotorula rubra was susceptible on 0.12-0.5 mg/ml and Saccharomyces cerevisiae on 0.25 mg/ml. Conclusions. Sage oil showed antifungal activity. The more susceptible to oil were the strains of Candida parapsilosis, Rhodotorula rubra and Saccharomyces cerevisiae. Oil was less active toward strains Candida glabrata, Candida tropicalis and Geotrichum candidum.

Piper diospyrifolium Kunth.: Chemical analysis and antimicrobial (intrinsic and combined) activities

The secular use of plants in popular medicine has emerged as a source for the discovery of new compounds capable of curing infections. Among microbial resistance to commercial drugs, species such as Piper diospyrifolium Kunth, which are used in popular therapy, are targets for pharmacological studies. With this in mind, antimicrobial experiments with the essential oil from the P. diospyrifolium (PDEO) species were performed and its constituents were elucidated. The oil compounds were identified by gas chromatography coupled to mass spectrometry (GC/MS). The broth microdilution method with colorimetric readings for bacterial tests (Escherichia coli and Staphylococcus aureus) and spectrophotometric readings for fungal tests (Candida albicans and Candida tropicalis), whose data were used to create a cell viability curve and calculate its IC50 against fungal cells, were used to determine the minimum inhibitory concentration of the oil and its combined action with commercial drugs. The oil's minimal fungicidal concentration and its action over fungal morphological transition were analyzed by subculture and microculture, respectively. Chemical analysis revealed Z-Carpacin, Pogostol and E-Caryophyllene as the most abundant compounds. Results from the intrinsic analysis were considered clinically irrelevant, however the oil presented a synergistic effect against multiresistant E. coli and S. aureus strains when associated with gentamicin, and against the standard and isolated C. tropicalis strains with fluconazole. A fungicidal effect was observed against the C. albicans isolate. Candida spp. hyphae inhibition was verified for all strains at the highest tested concentrations. The P. diospyrifolium essential oil presented a promising effect when associated with commercial drugs and against a fungal virulence factor. Thus, the oil presented active compounds which may help the development of new drugs, however, new studies are needed in order to clarify the oil's mechanism of action, as well as to identify its active constituents.

Antifungal and biofilm inhibitory effect of Cymbopogon citratus (lemongrass) essential oil on biofilm forming by Candida tropicalis isolates; an in vitro study.

Cymbopogon citratus (lemongrass) essential oil has been widely used as a traditional medicine and is well known for antimicrobial properties. Therefore, it might be a potent anti-infective and biofilm inhibitive against Candida tropicalis infections. Until now, no ideal coating or cleaning method based on an essential oil has been described to prevent biofilm formation of Candida strains on silicone rubber maxillofacial prostheses, voice prostheses and medical devices susceptible to C. tropicalis infections. To investigate the antifungal and biofilm inhibitory effects of Cymbopogon citratus oil. Clinical isolates of C. tropicalis biofilms on different biomaterials were used to study the inhibitory effect. The efficacy of Cymbopogon citratus, Cuminum cyminum, Citrus limon and Cinnamomum verum essential oils were compared on biofilm formation of three C. tropicalis isolates on 24 well polystyrene plates. C. citratus oil coated silicone rubber surfaces were prepared using hypromellose ointment as a vehicle. The antifungal tests to determine minimum inhibitory and minimum fungicidal concentrations were assessed by a microbroth dilution method and biofilm formation was determined by a crystal violet binding assay. C. tropicalis strains formed more biofilm on hydrophobic materials than on hydrophilic glass. C. citratus oil showed a high antifungal effect against all C. tropicalis strains. For comparison, C. limon oil and C. cyminum oil showed minor to no killing effect against the C. tropicalis strains. C. citratus oil had the lowest minimal inhibitory concentration of all essential oils tested and inhibited biofilm formation of all C. tropicalis strains. C. citratus oil coating on silicone rubber resulted in a 45-76% reduction in biofilm formation of all C. tropicalis strains. Cymbopogon citratus oil has good potential to be used as an antifungal and antibiofilm agent on silicone rubber prostheses and medical devices on which C. tropicalis biofilms pose a serious risk for skin infections and may cause a shorter lifespan of the prosthesis.

Xylitol production on sugarcane biomass hydrolysate by newly identified Candida tropicalis JA2 strain.

Xylitol is a building block for a variety of chemical commodities, besides being widely used as a sugar substitute in the food and pharmaceutical industries. The aim of this work was to develop a microbial process for xylitol production using sugarcane bagasse hydrolysate as substrate. In this context, 218 non-Saccharomyces yeast strains were screened by growth on steam-exploded sugarcane bagasse hydrolysate containing a high concentration of acetic acid (8.0g/L). Seven new Candida tropicalis strains were selected and identified, and their ability to produce xylitol on hydrolysate at low pH (4.6) under aerobic conditions was evaluated. The most efficient strain, designated C.tropicalis JA2, was capable of producing xylitol with a yield of 0.47g/g of consumed xylose. To improve xylitol production by C.tropicalis JA2, a series of experimental procedures were employed to optimize pH and temperature conditions, as well as nutrient source, and initial xylose and inoculum concentrations. C.tropicalis JA2 was able to produce 109.5g/L of xylitol with a yield of 0.86g/g of consumed xylose, and with a productivity of 2.81g·L·h, on sugarcane bagasse hydrolysate containing 8.0g/L acetic acid and177 g/L xylose, supplemented with 2.0g/L yeast nitrogen base and 4.0g/L urea. Thus, it was possible to identify a new C.tropicalis strain and to optimize the xylitol production process using sugarcane bagasse hydrolysate as a substrate. The xylitol yield on biomass hydrolysate containing a high concentration of acetic acidobtained in here is among the best reported in the literature.

Anticandidal activities of lactic acid bacteria isolated from the vagina

Background/aimLactic acid bacteria prevent the overgrowth of pathogenic agents and opportunistic pathogens in the vagina. Moreover, lactic acid bacteria contribute to the preservation of vaginal microbiota by producing antimicrobial agents. Previous studies showed that some lactic acid bacteria exhibited antimicrobial activity against Candida species causing yeast vaginosis as well as many bacterial pathogens.Materials and methods The antifungal activities of various lactic acid bacteria isolated from the vagina of healthy women on some Candida species isolated from the vagina were investigated by agar diffusion technique. Results Most of the lactic acid bacteria that belong to the species of Lactobacillus crispatus, L. fermentum, L. acidophilus, L. paracesei subsp. paracesei, L. pentosus, and L. plantarum exhibited antifungal activity in varying ratios against C. albicans, C. glabrata, and C. tropicalis strains isolated from the vagina.Conclusion The lactic acid bacteria are useful microorganisms associated with a variety of probiotic properties. In this sense, our lactic acid bacteria isolates with high antifungal activity may be promising candidates as probiotic microorganisms in the inhibition of vaginal candidiasis, which is one of the most prevalent problems, or in the protection against candidiasis. We will continue our studies in this area.

HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth (Lamiaceae) against Candida strains

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration (MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction (DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥ 4096 μg/mL against Candida albicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC50 of 2.60 and 2.53 μg/mL respectively, demonstrating significant synergistic effects against Candida strains. The modulator activity of DHm might be due to the presence of caffeic acid. Moreover, DHm and caffeic acid did not cause significant hemolytic effects, indicating that they present low cytotoxicity. Conclusions: These data indicate that DHm potentiates the activity of the fluconazole, without enhancement of the toxicity, encouraging further toxicological, pharmacological and phytochemical studies to provide consistent evidence of the potential of this plant to be used in drug development.

Effect of seasonality on chemical profile and antifungal activity of essential oil isolated from leaves Psidium salutare (Kunth) O. Berg.

Medicinal plants play a crucial role in the search for components that are capable of neutralizing the multiple mechanisms of fungal resistance. Psidium salutare (Kunth) O. Berg is a plant native to Brazil used as both food and traditional medicine to treat diseases and symptoms such as stomach ache and diarrhea, whose symptoms could be related to fungal infections from the genus Candida. The objective of this study was to investigate the influence of seasonal variability on the chemical composition of the Psidium salutare essential oil, its antifungal potential and its effect on the Candida albicans morphogenesis. The essential oils were collected in three different seasonal collection periods and isolated by the hydrodistillation process in a modified Clevenger apparatus with identification of the chemical composition determined by gas chromatography coupled to mass spectrometry (GC/MS). The antifungal assays were performed against Candida strains through the broth microdilution method to determine the minimum fungicidal concentration (MFC). Fungal growth was assessed by optical density reading and the Candida albicans dimorphic effect was evaluated by optical microscopy in microculture chambers. The chemical profile of the essential oils identified 40 substances in the different collection periods with γ-terpinene being the predominant constituent. The antifungal activity revealed an action against the C. albicans, C. krusei and C. tropicalis strains with an IC50 ranging from 345.5 to 2,754.2 µg/mL and a MFC higher than 1,024 µg/mL. When combined with essential oils at sub-inhibitory concentrations (MIC/16), fluconazole had its potentiated effect, i.e. a synergistic effect was observed in the combination of fluconazole with P.salutare oil against all Candida strains; however, for C. albicans, its effect was reinforced by the natural product in all the collection periods. The results show that the Psidium salutare oil affected the dimorphic transition capacity, significantly reducing the formation of hyphae and pseudohyphae in increasing concentrations. The results show that P. salutare oil exhibits a significant antifungal activity against three Candida species and that it can act in synergy with fluconazole. These results support the notion that this plant may have a potential use in pharmaceutical and preservative products.

Identification of Candida species by restriction enzyme analysis

Background/aim: The identification of Candida species isolated from clinical specimens provides information about antifungal susceptibility and sheds light on the choice of empirical treatment. In the present study, restriction enzyme analysis of C. albicans and non-albicans Candida species previously identified by conventional methods was done to evaluate the utility of restriction enzyme analysis for more rapid and reliable identification of Candida species. Materials and methods: A total of 146 Candida strains isolated from various clinical specimens and ATCC strains were included. PCR products were digested with MwoI for all species and with BslI for C. parapsilosis and C. tropicalis strains. Results: The strains were identified by conventional methods as 40 C. albicans, 27 C. parapsilosis, 26 C. tropicalis, 25 C. glabrata, 11 C. kefyr, 10 C. krusei, and 7 C. guilliermondii strains. Restriction digestion with MwoI was able to distinguish between five different species (C. albicans, C. krusei, C. guilliermondii, C. kefyr, and C. glabrata), while BslI digestion could distinguish between C. tropicalis and C. parapsilosis. Conclusion: Restriction enzyme analysis with MwoI and BslI can be used for the identification of Candida species in situations where rapid identification is necessary or conventional methods are problematic.

Virulence factors and determination of antifungal susceptibilities of Candida species isolated from palm wine and sorghum beer

Since it has been considered that Candida species in food or drinks may, following ingestion, enter the bloodstream and cause fungaemia, the presence of these yeast species in traditional alcoholic beverages may be of some clinical significance. Thus we attempted to assess virulence factors and antifungal susceptibility profile of Candida strains and other potential pathogenic yeasts isolated from palm wine and sorghum beer (tchapalo). Of the 23 yeast isolates from palm wine, phospholipase, esterase and haemolysin production was revealed amongst 69.6%, 65.2% and 100% isolates respectively with high activity belonging to Candida tropicalis strains. All the isolates were biofilm producers at variable degree but none showed proteinase activity. When the isolates were tested for their susceptibility to five antifungal agents, we found that ketoconazole (91.3%) followed by fluconazole (78.3%) and amphotericin B (73.9%) were the most potent agents. Of the 14 isolates from tchapalo, 57.1%, 87.5% and 57.1% exhibited phospholipase, haemolysin and esterase activity respectively. They did not also show proteinase activity while 87.5% produced biofilm. The majority of the isolates were susceptible to azoles (92.7%) and amphotericin B (85.3%) but they were 5-flucytosine resistant like palm wine strains.

Antifungal Activity of the Essential Oil of Cymbopogon citratus (DC) Stapf. An in vitro study.

Abstract: Aim. To evaluate the antifungal potential of the essential oil of Cymbopogon citratus by determining the Minimum Inhibitory Concentration (MIC) and the Minimum Fungicidal Concentration (MFC) for Candida albicans (ATCC 90029), Candida albicans (CBS 562), Candida tropicalis (ATCC 705) and Candida tropicalis strains (CBS 94), as well as to analyze the possible mechanism of action of the oil through the addition of sorbitol to the culture medium. Methods. For the MIC determination, inocula were previously adjusted through spectrophotometry and 100μL were added to the wells of plates already containing the culture medium and 100μL of the serial dilutions of the oil, incubating them in aerobiosis for 24 hours, with subsequent staining by 1% TCT. For the MFC, 50μL of the supernatant from the MIC assay wells were dripped onto Petri dishes and incubated in aerobiosis for 24 hours. Tests were performed in triplicate and data analysed by descriptive statistics. Results. It was determined that the MIC for C. albicans was 125 μg/mL while MIC for C. tropicalis was 250 μg/mL, with the essential oil presenting fungicidal effect for both analyzed yeasts. Conclusion. The essential oil of Cymbopogon citratus does not act at the cellular wall level and demonstrated an antimicrobial effect on Candida albicans and Candida tropicalis, therefore acting as a fungicide.

Action mechanism of naphthofuranquinones against fluconazole-resistant Candida tropicalis strains evidenced by proteomic analysis: The role of increased endogenous ROS

The increased incidence of candidemia in terciary hospitals worldwide and the cross-resistance frequency require the new therapeutic strategies development. Recently, our research group demonstrated three semi-synthetic naphthofuranquinones (NFQs) with a significant antifungal activity in a fluconazole-resistant (FLC) C. tropicalis strain. The current study aimed to investigate the action's preliminary mechanisms of NFQs by several standardized methods such as proteomic and flow cytometry analyzes, comet assay, immunohistochemistry and confocal microscopy evaluation. Our data showed C. tropicalis 24 h treated with all NFQs induced an expression's increase of proteins involved in the metabolic response to stress, energy metabolism, glycolysis, nucleosome assembly and translation process. Some aspects of proteomic analysis are in consonance with our flow cytometry analysis which indicated an augmentation of intracellular ROS, mitochondrial dysfunction and DNA strand breaks (neutral comet assay and γ-H2AX detection). In conclusion, our data highlights the great contribution of ROS as a key event, probably not the one, associated to anti-candida properties of studied NFQs.

Fern extracts potentiate fluconazole activity and inhibit morphological changes in Candida species

ObjectiveTo investigate the antifungal activity of the fern species Lygodium venustum (L. venustum) and Pityrogramma calomelanos (P. calomelanos) against Candida albicans and Candida tropicalis strains. MethodsThe microdilution method was used to evaluate the antifungal activity, as well as the modulating effects of ethanolic extracts of these plants in combination with fluconazole. The minimum inhibitory concentration (MIC), minimum fungicide concentration and morphological changes were also determined. ResultsThe extract obtained from L. venustum presented a MIC > 8192 μg/mL, while the extract obtained from and P. calomelanos presented a MIC = 8192 μg/mL, indicating that they present weak antifungal activity. However, combination of the extracts with Fluconazole potentiated the antifungal activity of this drug. At different experimental conditions, such as concentration of the extract and type of strain, the extracts inhibited hyphae and pseudohyphae formation, indicating that these fern species can affect the morphology of the fungi. ConclusionsThe extracts obtained from the fern species L. venustum and P. calomelanos dose not present significant antifungal activity. However, P. calomelanos potentiates the activity of fluconazole and both extracts inhibits the morphological changes in Candida species, indicating that they have potential pharmacological activity as modulators of fungal biology. Therefore, novel studies are required to characterize the interference of these extracts in the virulence and pathogenicity of Candida species as well as the potential of fern species to treat fungal infections.

Population structure and molecular genetic characterization of clinical Candida tropicalis isolates from a tertiary-care hospital in Kuwait reveal infections with unique strains.

Candida tropicalis is a frequently isolated yeast species causing bloodstream, urinary tract and other infections particularly in patients admitted to intensive care units (ICUs) and those requiring prolonged urinary catheterization (UC) or receiving broad-spectrum antibiotics (BSA). This study investigated clinical characteristics and genetic relatedness among C. tropicalis strains isolated from patients at Al-Amiri Hospital in Kuwait. C. tropicalis strains (n = 63) isolated from blood, genito-urinary, respiratory (RT) and digestive (GIT) tracts and wound sites from 54 patients were used. All isolates were phenotypically identified and tested against six antifungal drugs by using Vitek 2 system. Molecular identification was performed by PCR amplification of rDNA. Fingerprinting was achieved by 6-loci-based multilocus sequence typing (MLST) and data were analyzed by BioNumerics software for phylogenetic relationships. Patients mean age was >65 years and >20% patients were hospitalized in ICUs. Most patients had underlying conditions that included UC, BSA, diabetes and RT/GIT abnormalities. Most candiduria cases had UC, ureteric stent or suprapubic catheters. All isolates were identified as C. tropicalis by Vitek 2 and by species-specific PCR. Sixty-two isolates were susceptible to all tested antifungal drugs. MLST identified 59 diploid sequence types (DSTs) including 54 newly-identified DSTs. C. tropicalis isolates from multiple sites of same patient usually belonged to different DSTs. Interestingly, 56 of 57 isolates from 48 patients belonged to unique genotypes. Only six isolates from six patients belonged to three DSTs (clusters), however, C. tropicalis strains in each cluster were isolated >3 months apart. Our data show diverse origins of C. tropicalis infections in Kuwait as most isolates were unique strains. There was no obvious correlation between cluster isolates with time of isolation and/or hospital ward of their origin. This study presents the first MLST analysis of C. tropicalis isolates from Middle East and may be useful for studying genetic relationships among global C. tropicalis strains.

Notable Increasing Trend in Azole Non-susceptible Candida tropicalis Causing Invasive Candidiasis in China (August 2009 to July 2014): Molecular Epidemiology and Clinical Azole Consumption.

Objectives: To report the notable increasing trends of C. tropicalis antifungal resistance in the past 5 years, and explore molecular epidemiology, and the relationship between clinical azoles consumption and increased resistance rate.Methods: Between August 2009 and July 2014, 507 non-duplicated C. tropicalis isolates causing invasive candidiasis were collected from 10 hospitals in China. The in vitro antifungal susceptibility of nine common agents was determined by Sensititre YeastOne™ using current available species-specific clinical breakpoint (CBPs) or epidemiological cut-off values (ECVs). A high discriminatory three-locus (ctm1, ctm3, and ctm24) microsatellite scheme was used for typing of all isolates collected. Clinical consumption of fluconazole and voriconazole was obtained and the Defined Daily Dose measurement units were assigned to the data.Results: Overall, 23.1 and 20.7% of isolates were non-susceptible to fluconazole and voriconazole, respectively. And over 5 years, the non-susceptible rate of C. tropicalis isolates to fluconazole and voriconazole continuously increased from 11.2 to 42.7% for fluconazole (P < 0.001), and from 10.4 to 39.1% for voriconazole (P < 0.001). Four genotype clusters were observed to be associated with fluconazole non-susceptible phenotype. However, the increase in azole non-susceptible rate didn't correlate with clinical azole consumption.Conclusions: The rapid emergence of azole resistant C. tropicalis strains in China is worrying, and continuous surveillance is warranted and if the trend persists, empirical therapeutic strategies for C. tropicalis invasive infections should be modified.

Antifungal effect of 7-hydroxycitronellal against C. Tropicalis strains: an in vitro approach

The fungi of the genus Candida play a relevant role in the emergence of oral infections, and are increasingly more frequent the cases of infections by non- albicans strains. In light of this context and the need for new alternatives to the antimicrobial therapy, the monoterpene [7-hidroxicitronelal] (7-HO) was evaluated for its antifungal effects. For the obtainment of the MIC and MFC values the broth microdilution method was used. The MICand the MFC of this monoterpene for 60% of the tested strains was of 256µg/mL and 512µg/mL respectively. Furthermore the standard antifungal Nystatin (100 UI/mL) was used as positive control for the inhibition of fungal growth. Therefore, were used 4 clinical strains of the species tropicalis (LM 06, LM 14, LM 31 and LM 36) and a standard strain ( C. tropicalis ATCC 13803), originated from the Mycology collection of the Mycology Laboratory (LM) of the Health Sciences Center (CCS) of the Federal University of Paraiba (UFPB). The results obtained in this study showed fungicide activity of the compound (7-OH) against the strains of Candida tropicalis .

Evaluation of some lignocellulosic byproducts of food industry for microbial xylitol production by Candida tropicalis.

Some lignocellulosic food byproducts such as potato peels, wheat bran, barley bran and chestnut shells were evaluated as potential sources of xylose for microbial xylitol production by yeasts. Potential yeast strains were selected after screening xylitol production of some indigenous yeasts in a defined fermentation medium. Candida tropicalis strains gave the highest results with 83.28 and 54.07 g/L xylitol production from 100 g/L xylose. Lignocellulosic materials were exposed to acid hydrolysis at different conditions. Chestnut shells gave the highest xylose yield and the hydrolysate of chestnut shells was used in further experiments in which xylitol productions of two potential C. tropicalis strains were investigated. Combined detoxification method including evaporation, overliming and activated charcoal with the use of threefold concentration and also yeast extract supplementation suggested to be efficient for both growth and product formation in chestnut shell hydrolysate in which 40 % xylitol yield was obtained. It was concluded that detoxified and fortified chestnut shell hydrolysate could be a potential medium for xylitol production.

Candida tropicalis biofilm inhibition by ZnO nanoparticles and EDTA

ObjectiveBiofilm of Candida tropicalis denote as a complex cellular congregation with major implication in pathogenesis. This lifestyle of fungus as a biofilm can inhibit immune system and antifungal therapy in treatment of infectious disease especially medical device associated chronic disease. In this study effects of Zinc Oxide (ZnO) nanoparticles and EDTA were evaluated on C. tropicalis biofilm by using different techniques. ZnO nanoparticles were synthesized from Egg albumin. DesignTo assay the formation of biofilm of yeast cells like Fluconazole-susceptible C. tropicalis (ATCC 13,803) and fluconazole-resistant standard strains of C. tropicalis (ATCC 750) were grown in 24 well plates and antifungal effect of ZnO and EDTA were evaluated on C. tropicalis biofilm using ATP bioluminescence and tetrasodium salt (XTT) reduction assays. ResultsSynthesized ZnO NPs and EDTA had effective antifungal properties at the concentration of 5.2, 8.6μg/ml for Fluconazole susceptible strain and 5.42, 10.8μg/ml Fluconazole resistant strains of C. tropicalis biofilms compared to fluconazole drug. ConclusionIn present study we conclude, ZnO considered as a new agent in field of prevention C. tropicalis biofilms especially biofilms formed surface of medical device.

The Role of UPC2 Gene in Azole-Resistant Candida tropicalis.

Azole resistance of Candida tropicalis has, in recent years, become a serious issue in hospitals; however, there is limited knowledge of the mechanisms underlying this resistance. We have previously demonstrated that ERG11 plays a vital role in azole resistance in C. tropicalis. Here, we describe the expression and sequence variation of UPC2, which encodes a transcription factor of ERG11. Quantitative real-time RT-PCR showed that 31 azole-resistant C. tropicalis strains significantly overexpressed UPC2. Those isolates resistant to all three azole antifungals upregulated UPC2 expression to a greater degree than those resistant to only fluconazole or itraconazole. The UPC2 promoter contains mutations -118T-G and -155G-A in azole-resistant strains of C. tropicalis. Meanwhile, the mutation G392E was also detected twice in UPC2 gene in azole-resistant C. tropicalis and was demonstrated to mediate azole antifungal susceptibility by using Saccharomyces cerevisiae as an expression host, particularly for fluconazole and itraconazole.

Distinct Expression Levels of ALS, LIP, and SAP Genes in Candida tropicalis with Diverse Virulent Activities.

Candia tropicalis is an increasingly important human pathogen, causing nosocomial fungemia among patients with neutropenia or malignancy. However, limited research has been published concerning its pathogenicity. Based on the phenotypes of C. tropicalis in our previous study, we selected nine representative strains with different activities of virulence factors (adhesion, biofilm formation, secreted aspartic proteinases, and hemolysins), and one reference strain, ATCC750. The present study aimed to investigate the filamentation ability, the expression of virulence genes (ALST1-3, LIP1, LIP4, and SAPT1-4) and the cell damage of C. tropicalis strains with diverse virulences. C. tropicalis exhibited strain-dependent filamentation ability, which was positively correlated with biofilm formation. Reverse transcriptase PCR analysis showed that the ALST3 and SAPT3 genes had the highest expression in their corresponding genes for most C. tropicalis. The expressions of virulence genes, except ALST3 on polystyrene, were upregulated compared with growth in the planktonic and on human urinary bladder epithelial cell line (TCC-SUP) surface. Clustering analysis of virulence genes showed that isolates had a high biofilm forming ability on polystyrene formed a group. Lactate dehydrogenase assays showed that the cell damage induced by C. tropicalis markedly increased with longer infection time (24 and 48 h). Strain FXCT01, isolated from blood, caused the most serious cell damage; while ZRCT52, which had no filamentation ability, caused the least cell damage. Correlation analysis demonstrated significant correlation existed between adhesion on epithelial cells or the expression of ALST2-3 and cell damage. Overall, our results supported the view that adhesion and filamentation may play significant roles in the cell damage caused by C. tropicalis.