Inhibited Germ Tube Formation Research Articles

Studies on the antifungal effects of Hinokitiol on Candida albicans: inhibition of germ tube formation and synergistic pharmacological effects of miconazole.

One of the goals of oral healthcare management is to manage dry mouth. Thus, moisturizers containing antimicrobial ingredients, such as hinokitiol (HT), are applied to the oral mucosa after oral care. In this study, we investigated the preventive effect of HT against the growth of Candida albicans (C. al) and its synergistic effect when combined with miconazole (MCZ), an oral treatment for candidiasis. As the concentration of HT increased, the length and percentage of germ tubes (GT) decreased. Larger inhibition circles were observed for MCZ concentrations of 2.0 and 4.0μg/disc compared to the HT medium without HT. The increased inhibitory effect was observed in both aerobic and anaerobic cultures. This suggests that the production of reactive oxygen species (ROS) by C. al cells increased with the combination of HT and MCZ. The length and percentage of GT increased, whereas the amount of ROS decreased when ROS scavengers were used in combination with the drug. HT led to morphological changes that inhibited the GT associated with pathogenic C. al, exhibited a complementary action against MCZ, and showed a possible association with hydrogen peroxide and superhydroxy anion radicals. These effects suggest that HT is a promising candidate for inhibiting C. al. In conclusion, HT demonstrated a prophylactic effect by inhibiting C. al and a synergistic effect with MCZ, a drug used to treat oral candidiasis. HT may also be useful for suppressing the onset and reducing the severity of oral candidiasis.

Crinum latifolium mediated biosynthesis of gold nanoparticles and their anticandidal, antibiofilm and antivirulence activity

Candida spp. is one of the most common opportunistic human fungal pathogens, responsible for 90–100% of mucosal infections. Germ tube formation, hyphal morphogenesis, the production of tissue-damaging extracellular enzymes, and drug-resistant biofilm formation contribute to their pathogenicity, which can lead to systemic infections in the worst scenarios. Thus, there is an urgent need to discover new therapeutic agents to overcome the above virulence factors. Therefore, we aimed to prepare Crinum latifolium leaves-mediated biosynthesis of gold nanoparticles (AuNPs) that was characterized by various sophisticated techniques, and further their antifungal, antibiofilm, and anti-virulence activities was investigated. The AuNPs show a zone of inhibition between 19 and 22 mm for test strains of Candida spp. at 1000 µg/ml, whereas the MIC values were ranged from 250 to 500 µg/ml. AuNPs inhibit germ tube formation in C. albicans by 93.3% at 50 µg/ml. Furthermore, exposure to AuNPs significantly reduced the secretions of phospholipase, proteinase, hemolysin, esterase, and lipase. Confocal laser scanning microscopy (CLSM) investigation shows that 25 µg/ml of AuNPs significantly inhibit colonization and biofilm formation. Lastly, the interaction of Candidal cells with AuNPs revealed, ultrastructural changes in the cell wall and cell membranes as visualized by a transmission electron microscope (TEM).

Antifungal Activity of Mexican Propolis on Clinical Isolates of Candida Species.

Infections caused by micro-organisms of the genus Candida are becoming a growing health problem worldwide. These fungi are opportunistic commensals that can produce infections—clinically known as candidiasis—in immunocompromised individuals. The indiscriminate use of different anti-fungal treatments has triggered the resistance of Candida species to currently used therapies. In this sense, propolis has been shown to have potent antimicrobial properties and thus can be used as an approach for the inhibition of Candida species. Therefore, this work aims to evaluate the anti-Candida effects of a propolis extract obtained from the north of Mexico on clinical isolates of Candida species. Candida species were specifically identified from oral lesions, and both the qualitative and quantitative anti-Candida effects of the Mexican propolis were evaluated, as well as its inhibitory effect on C. albicans isolate’s germ tube growth and chemical composition. Three Candida species were identified, and our results indicated that the inhibition halos of the propolis ranged from 7.6 to 21.43 mm, while that of the MFC and FC50 ranged from 0.312 to 1.25 and 0.014 to 0.244 mg/mL, respectively. Moreover, the propolis was found to inhibit germ tube formation (IC50 ranging from 0.030 to 1.291 mg/mL). Chemical composition analysis indicated the presence of flavonoids, including pinocembrin, baicalein, pinobanksin chalcone, rhamnetin, and biochanin A, in the Mexican propolis extract. In summary, our work shows that Mexican propolis presents significant anti-Candida effects related to its chemical composition, and also inhibits germ tube growth. Other Candida species virulence factors should be investigated in future research in order to determine the mechanisms associated with antifungal effects against them.

Activity of poly(methacrylic acid)-silver nanoparticles on fluconazole-resistant Candida albicans strains: Synergistic and cytotoxic effects.

To synthesize and evaluate the antifungal activity of poly(methacrylic acid)-silver nanoparticles (PMAA-AgNPs) against nine Candida albicans isolated from clinical specimens. The effects of PMAA-AgNPs-fluconazole combination was analysed by checkerboard methodology. The synergistic potential of PMAA-AgNPs-fluconazole was determined by the fractional inhibitory concentration index (FICI). The inhibition of germ tube formation and the determination of PMAA-AgNPs cytotoxicity were also performed. All C. albicans strains were susceptible to PMAA-AgNPs and resistant to fluconazole. PMAA-AgNPs at subinhibitory concentrations restored the susceptibility of resistant C. albicans to fluconazole, whose FICI ranged from 0.3 to 0.5. The synergistic interaction of the combination was observed in eight of nine strains. The PMAA-AgNPs-fluconazole combination was also able to inhibit the germ tube formation. PMAA-AgNPs showed a dose-dependent decrease in viability for cells tested, with 50% cytotoxic concentration (CC50 ) values of 6.5, 4.9 and 6.8μg ml-1 for macrophages, fibroblasts and Vero cells, respectively. This study demonstrated that, in general, PMAA-AgNPs acts synergistically in combination with fluconazole, inhibiting fluconazole-resistant C. albicans strains. PMAA-AgNPs-fluconazole combination was also able to inhibit germ tube formation, an important virulence factor. Inhibitory effect of PMAA-AgNPs alone or in combination was higher in C. albicans than in mammalian cells. This study shows the potential of PMAA-AgNPs combined with fluconazole to inhibit fluconazole-resistant C. albicans strains.

The novel antifungal agent AB-22 displays in vitro activity against hyphal growth and biofilm formation in Candida albicans and potency for treating systemic candidiasis.

Systemic candidiasis, which is mainly caused by Candida albicans, is a serious acute fungal infection in the clinical setting. In a previous study, we reported that compound 22h (designated as AB-22 in this study), a vinyl sulfate compound, is a fast-acting fungicidal agent against a broad spectrum of fungal pathogens. In this study, we aimed to further analyze the in vitro and in vivo efficacy of AB-22 against filamentation, biofilm formation, and virulence of C. albicans. Under in vitro hyphal growth-inducing condition, AB-22 effectively inhibited germ tube formation and hyphal growth, which are required for the initiation of biofilm formation. Indeed, AB-22 significantly suppressed C. albicans biofilm formation in a dose-dependent manner. Moreover, AB-22 treatment inhibited the normal induction of ALS3, HWP1, and ECE1, which are all required for hyphal transition in C. albicans. Furthermore, AB-22 treatment increased the survival of mice systemically infected with C. albicans. In conclusion, in addition to its fungicidal activity, AB-22 inhibits filamentation and biofilm formation in C. albicans, which could collectively contribute to its potent in vivo efficacy against systemic candidiasis.

Inhibition of Candida albicans in vivo and in vitro by antimicrobial peptides chromogranin A-N12 through microRNA-155/suppressor of cytokine signaling 1 axis

ABSTRACT Antimicrobial peptides (AMPs) have proven to inhibit a variety of pathogens. Chromogranin A-N12 (CGA-N12) is a kind of AMP, and it is characterized by stable structure, high anti-Candida activity, and good safety. However, it remains unclear whether CGA-N12 could effectively inhibit the growth of Candida albicans (C. albicans). Colony forming assays were used to measure minimal inhibitory concentration (MIC), minimal fungicidal concentration (MFC), and time-kill curve. Disseminated C. albicans rabbit model was established to investigate the influence of CGA-N12 on histological damage. The protein and mRNA levels of suppressor of cytokine signaling 1 (SOCS1) after treatment were investigated. The MIC and MFC of CGA-N12 against C. albicans was 6 mg/mL. CGA-N12 considerably inhibited germ tube formation of C. albicans. The fungal load in the tissues and inflammatory factors in the serum were suppressed by CGA-N12. CGA-N12 significantly reduced the histological changes caused by C. albicans, and the protein and mRNA levels of SOCS1 were markedly inhibited. The inhibition effect of CGA-N12 on C. albicans and significant improvement of histological damage by CGA-N12 through microRNA-155/SOCS1 axis were proved in this study. This study proposes a novel therapeutic strategy for the treatment and prevention of C. albicans. Abbreviations: AMPs: Antimicrobial peptides; MIC: Minimal inhibitory concentration; MFC: Minimal fungicidal concentration; AIDS: Acquired immune deficiency syndrome; PBS: Phosphate buffer saline; FBS: Fetal bovine serum; ROS: Reactive oxygen species; CFU: Colony formation unit; CGA: Chromogranin A; SOCS1: Suppressor of cytokine signaling 1; SDA: Sabouraud Dextrose Agar; GRAVY: Grand average of hydropathicity; C. parapsilosis: Candida parapsilosis; C. albicans: Candida albicans

Inhibition of Germ Tube Formation in Mucor & Candida by JRK’s DCOD

JRK’s DOCD is a proprietary siddha medicine, indicated for prevention of co-morbidities of diabetes. Earlier studies on the product has proven its effect in boosting the phagocyte mediated immunity. In the current study the ability of the product in preventing pathogenicity of fungi is evaluated. Germ tube formation is proven to increase the pathogenicity of the fungi than the spores alone. For this different species of fungi i.e., Candida albicans, 2 isolates of Candida dubelensis and three isolates of Mucor and 1 isolate of Rhizopus are taken. These fungal cultures when treated with the JRK’s DCOD, there is an inhibition of germ tube formation compared to control. Complete study details are presented in the paper.

Synergistic potential of essential oils with antibiotics to combat fungal pathogens: Present status and future perspectives.

The steady rise in the emergence of antibiotic-resistant fungal pathogens has rendered most of the clinical antibiotics available in the market to be ineffective. Therefore, alternative strategies are required to tackle drug-resistant fungal infections. An effective solution is to combine the available antibiotics with adjuvants such as phytochemicals or essential oils to enhance the efficacy and activity of antibiotics. The present review aims to summarize the studies on synergistic combinations of essential oils and anti-fungal antibiotics. The current findings, methods used for measuring synergistic effects, possible mechanisms of synergism, and future perspectives for developing synergistic EO-antibiotic therapeutic formulations are discussed in this study. Several essential oils exhibit synergistic effect in combination with antibiotics against human fungal pathogens such as Candida albicans. The possible mechanisms of synergy exhibited by essential oil- antibiotic combinations in fungi include disruption of cell wall structure/ ergosterol biosynthesis pathway, enhanced transdermal penetration of antibiotics, alterations in membrane permeability, intracellular leakage of cellular contents, inhibition of germ tube formation or fungal biofilm formation, and competition for a primary target. Synergistic combination of essential oils and antibiotics can prove to be a valid and pragmatic alternative to develop drugs with increased drug-efficacy, and low toxicity.

Evaluation of the antifungal photodynamic activity of Thymophylla pentachaeta extracts against Candida albicans and its virulence factors

BackgroundCandida albicans is one of the most common causative of opportunistic infections. Treatment of candidiasis is challenging considering the few antifungal drugs available and the increase in resistance. Antimicrobial photodynamic therapy (aPDT) is a recently developed therapeutic option that combines a non-toxic photosensitizer (PS) and light to kill the microbial pathogens. Targeting virulence, defined as the ability of a pathogen to cause overt disease, represents another attractive target for the development of novel antifungal agents. Thymophylla pentachaeta (DC.) Small var. belenidium (DC.) is an endemic plant from Argentina in which the presence of thiophenes, biologically active compounds whose antifungal activity is enhanced by irradiation with Ultraviolet A (UVA), have been already described. PurposeThe purpose of this study was to evaluate the photodynamic antifungal activity of hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc) and methanol (MeOH) extracts from T. pentachaeta var. belenidium and their inhibitory effects on C. albicans virulence factors as well as biofilm formation and eradication. Study Design/MethodsAntifungal photodynamic activity of Hex, DCM, EtOAc and MeOH extracts from different parts of the plant were assessed with the microbroth dilution, bioautography and the time-kill assays, under light and darkness conditions. The capacities of the most active extracts of inhibiting Candida virulence factors (adherence to epithelial cells, germ tube and pseudomycelium formation and hydrolytic enzyme secretion) were assessed. In addition, the activity against biofilm formation and eradication has been investigated by reaction with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) that quantifies living cells in these structures. ResultsHex and DCM extracts from T. pentachaeta roots exhibited high photodynamic antifungal activity against C. albicans [Minimal fungicide concentrations (MFCs)= 7.8 µg/ml] under UVA light irradiation. Chemical analysis of active extracts (Hex and DCM from roots) revealed the presence of photoactive thiophenes. Both extracts generate reactive oxygen species through type I and II mechanisms. These extracts, at sub-inhibitory concentrations, under light conditions decreased the adherence of C. albicans to Buccal Epithelial Cells (BEC), inhibited germ tube formation and reduced esterase production. Finally, they demonstrated activity against preformed biofilms submitted to irradiation (MFCs= 3.91 µg/ml and 15.63 µg/ml for Hex and DCM extracts, respectively). ConclusionTaking together, results demonstrated the strong photodynamic effects of T. pentachaeta root extracts under UVA irradiation, making them valuable alternatives to the already established antifungal drugs against C. albicans.

Non-antifungal drugs inhibit growth, morphogenesis and biofilm formation in Candida albicans.

The increased resistance/tolerance of Candida infections to antimicrobial treatment can be attributed to biofilm-associated cells. A way to overcome this situation is to re-purpose non-anti-fungal drugs that could be active against fungi. We have explored the potential of a small library of eighteen non-antifungal drugs used in different human diseases. Candida albicans was cultured in the presence and absence of different concentrations of these drugs. Subsequently, inhibition of growth, germ tube formation, adhesion, and biofilm development were studied. Out of eighteen drug molecules, six showed a reduction in planktonic and biofilm growth in a dose-dependent manner and three drugs inhibited germ tube formation. This study shows the potential of non-antifungal drugs for the development of new anti-Candida agents.

Dodonaea viscosa var angustifolia derived 5,6,8-trihydroxy-7,4′ dimethoxy flavone inhibits ergosterol synthesis and the production of hyphae and biofilm in Candida albicans

Ethnopharmacological relevanceCandida albicans is developing resistance to existing drugs increasing morbidity and mortality, which elevates an immediate need to explore new antifungal agents. Phytochemicals are an excellent source of therapeutic agents. We previously reported the antifungal activity of the crude extract of Dodonaea viscosa var. angustifolia Jacq. (DVA) from which a beneficial compound flavone: 5,6,8-trihydroxy-7,4′ dimethoxy flavone (5,6,8-trihydroxy-7-methoxy-2-(4-methoxyphenyl)-4H-chromen-4-one) abbreviated as TMMC, was extracted. Aim of the studyThe present study evaluated the effect of a TMMC subfraction on biofilms, membrane stability, ergosterol biosynthesis and germ tube (GT) formation in Candida albicans. Materials and methodsExtracts were prepared and fractionated to obtain purified TMMC. Minimum inhibitory concentrations of TMMC were obtained and subinhibitory concentrations were selected for further studies. Confocal laser scanning microscopy (CLSM) was performed to assess the effect of TMMC on membrane permeability and sterol deposition using propidium iodide (PI) and filipin stains, respectively. ResultsMinimum inhibitory concentrations (MIC) and Minimum Fungicidal concentrations (MFC) of TMMC were 0.39 mg/mL and 1.56 mg/mL, respectively. TMMC inhibited biofilm formation and damaged mature biofilms at 0.39 mg/mL and 1.56 mg/mL, respectively. CLSM further confirmed the disruption and architectural changes in biofilms following treatment with TMMC. TMMC also inhibited GT formation and ergosterol biosynthesis in a concentration dependent manner, which was further confirmed by varying sterol distribution and membrane disruption in treated and untreated cells. ConclusionsWith the multiple targets at different concentrations, TMMC warrants its potential use as antifungal drug against C. albicans. However further studies using animal models and more mechanistic approaches will be required.

Anticandidal activity of biosynthesized silver nanoparticles: effect on growth, cell morphology, and key virulence attributes of Candida species.

Purpose: The pathogenicity in Candida spp was attributed by several virulence factors such as production of tissue damaging extracellular enzymes, germ tube formation, hyphal morphogenesis and establishment of drug resistant biofilm. The objective of present study was to investigate the effects of silver nanoparticles (AgNPs) on growth, cell morphology and key virulence attributes of Candida species.Methods: AgNPs were synthesized by the using seed extract of Syzygium cumini (Sc), and were characterized by UV-Vis spectrophotometer, Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), and transmission electron microscopy (TEM). ScAgNPs were used to evaluate their antifungal and antibacterial activity as well as their potent inhibitory effects on germ tube and biofilm formation and extracellular enzymes viz. phospholipases, proteinases, lipases and hemolysin secreted by Candida spp.Results: The MICs values of ScAgNPs were ranged from 0.125-0.250 mg/ml, whereas the MBCs and MFCs were 0.250 and 0.500 mg/ml, respectively. ScAgNPs significantly inhibit the production of phospholipases by 82.2, 75.7, 78.7, 62.5, and 65.8%; proteinases by 82.0, 72.0, 77.5, 67.0, and 83.7%; lipase by 69.4, 58.8, 60.0, 42.9, and 65.0%; and hemolysin by 62.8, 69.7, 67.2, 73.1, and 70.2% in C. albicans, C. tropicalis, C. dubliniensis, C. parapsilosis and C. krusei, respectively, at 500 μg/ml. ScAgNPs inhibit germ tube formation in C. albicans up to 97.1% at 0.25 mg/ml. LIVE/DEAD staining results showed that ScAgNPs almost completely inhibit biofilm formation in C. albicans. TEM analysis shows that ScAgNPs not only anchored onto the cell surface but also penetrated and accumulated in the cytoplasm that causes severe damage to the cell wall and cytoplasmic membrane.Conclusion: To summarize, the biosynthesized ScAgNPs strongly suppressed the multiplication, germ tube and biofilm formation and most importantly secretion of hydrolytic enzymes (viz. phospholipases, proteinases, lipases and hemolysin) by Candia spp. The present research work open several avenues of further study, such as to explore the molecular mechanism of inhibition of germ tubes and biofilm formation and suppression of production of various hydrolytic enzymes by Candida spp.

Antifungal activity of some Sternbergia taxa: effects on germ tube and biofilm formation

Natural products are rapidly becoming the primary sources of novel antimicrobial agents, as resistance to existing antimicrobial agents is increasing. Apart from determining the antimicrobial activity of natural products, it is also important to understand their effects on the virulence factors of microorganisms. This study aimed to determine the antimicrobial activity of Sternbergia species prevalent in Turkey and investigate their role in the inhibition of germination tube and biofilm formation, both of which are known to be important virulence factors of Candida albicans. The antimicrobial activities of the plant extracts were evaluated using bore-plate and broth microdilution method. The extracts’ capacity to inhibit the formation of the germ-tube was also evaluated. The findings of our study revealed that Sternbergia lutea, Sternbergia vernalis possessed antimicrobial activities, with MIC values ranging between 0.048 mg/mL and 0.39 mg/mL. The highest antimicrobial activity was observed against Candida dubliniensis (0.048 mg/mL). While evaluating the inhibition of fungal germination activities, S. vernalis extract (at a concentration of 0.09 mg/mL) was found to be the most effective against C. albicans ATCC 90028 strain. The results also indicated that S. vernalis extracts at sub-MIC levels inhibited germ tube formation and modulated the tail-length of germinated cells, both of which are important virulence factors of C. albicans. Furthermore, the inhibition of biofilm-formation was also investigated, and it was found that two Sternbergia spp. extracts at or below MIC levels inhibited biofilm formation.

Effects of Ultraviolet Radiation (UVA+UVB) on Germination of Carpospores of the Red Macroalga Pyropia acanthophora var. brasiliensis (Rhodophyta, Bangiales): Morphological Changes.

Carpospores of Pyropia acanthophora var. brasiliensis are dispersion and reproduction units responsible for giving rise to the diploid filamentous structure of this alga's life cycle. The present study assesses the anthropogenic impact of ultraviolet radiation (UVR) on morphology and ultrastructure, spore viability, autofluorescence of chloroplasts and the amount of intensity of ROS during the germination of carpospores. Carpospores were cultivated at 24±1°C, 40±10μmolphotonsm-2 s-1 with photoperiod of 12h and exposed to UVAR+UVBR for 3h a day for 2days with a daily dose of 5.05Jcm-2 for UVAR and 0.095Jcm-2 for UVBR. Samples were cultured for another five days exposed only to PAR in order to confirm their viability after the initial 2-day exposure. Carpospores showed significant sensitivity to UVR exposure after only 48h, including changes in developmental rate, overall morphology, cell organization and chloroplast autofluorescence. UVR exposure inhibited germ tube formation in carpospores, which were mostly nonviable and/or altered, showing retracted cytoplasm and disorganized cytoplasmic content. Even in the absence of UVR exposure, carpospores remained collapsed, indicating irreversible damage. It can be concluded that UVR is a limiting factor for the development of P.acanthophora.

Satureja khuzistanica Jamzad essential oil and its anti-candidal activities against clinical isolates of Candida albicans isolated from women with candidiasis

Satureja khuzistanica Jamzad is known as antiseptic and analgesic agent in folk medicine. The aim of this investigation was to evaluate the anti-candidal activity of S. khuzistanica aerial parts essential oil against clinical isolates of Candida albicans, which were isolated from women with chronic recurrent candidiasis. For this purpose, the chemical composition of hydro-distilled essential oil was determined by GC and GC-MS analysis. Then, the anti-candidal activity of essential oil and its main component (carvacrol) were determined. Carvacrol (94.1%) was the main component of essential oil, followed by β-bisabolene, p-cymene and γ-terpinene. S. khuzistanica essential oil had strong anti-candidal activity against clinical isolates of C. albicans via inhibition of germ tube formation and induction the huge punctures in the cytoplasmic structures. The cell membranes were intact in presence of essential oil or carvacrol. S. khuzistanica essential oil as the main source of carvacrol can be used for treatment of C. albicans related infections.

Ocimum tenuiflorum L. and Ocimum basilicum L., two spices of Lamiaceae family with bioactive essential oils

The present study provides new insights to the antifungal mechanism of action of the essential oils of Ocimum tenuiflorum L. and Ocimum basilicum L., namely inhibition of germ tube formation, inhibition of biofilm formation and preformed biofilm disruption. The essential oils were characterized by GC and GC–MS. The major compounds were methyl eugenol (84.7%) and β-caryophyllene (7.4%) for O. tenuiflorum and linalool (35.1%), eugenol (20.7%) and 1,8-cineole (9.9%) for O. basilicum. The essential oil from O. tenuiflorum showed a more preeminent effect against C. neoformans (0.16 μL/mL) and dermatophytes (0.32 μL/mL). The effect on the germ tube formation of both essential oils was described here for the first time showing that O. tenuiflorum decreases germ tube formation by more than 50% at values four times lower than MIC (Minimal Inhibitory Concentration) while O. basilicum is able to decrease at values eight times lower than MIC. Furthermore, O. basilicum showed a more preeminent effect both in inhibition of C. albicans biofilm formation as well as in disruption of preformed biofilm. The activity of all major compounds was also determined, and their activity was in general similar to that of the essential oils thus suggesting that those are the main active compounds. Overall, this study highlights the antifungal activity of two widely used spices and complies with the antifungal uses described in folk medicine. In addition, it shows that both essential oils are able to inhibit virulence factors of C. albicans associated with resistance to treatment and relapse cases. Both species are of industrial interest as shown by their use on food and cosmetic industries which was reinforced by the results presented herein.

The efficacy of gaseous ozone against different forms of Candida albicans

Background and Purpose:Ozone is an inorganic molecule with effective antimicrobial properties. Clinical treatment of ozonated water was used for the elimination of Candida albicans, Enterococcus faecalis, endotoxins, and biofilms from root canals. In addition, its therapeutic effects for tinea pedis, ulcers, and leishmaniasis were investigated. The purpose of the present study was to evaluate the fungicidal effects of ozone on different forms of C. albicans. In addition, antifungal susceptibility profile of strains was assessed before and after exposure to ozone.Materials and Methods:Fifty strains of C. albicans were exposed to gaseous ozone at different times. Furthermore, biofilm formation and germ tube production were evaluated when yeast suspensions were exposed to ozone. In addition, antifungal susceptibility of ozone resistant colonies was investiagted as compared to controls.Results:Ozone was highly effective in killing C. albicans in yeast form and inhibition of germ tube formation during 210 and 180 s, respectively. Although with increasing exposure time biofilm production was considerably decreased, resistance to ozone was much higher among vaginal and nail isolates even after 60 min. All the strains were sensitive to fluconazole, caspofungin, and terbinafine pre- and post-ozone exposure. Resistance to amphotericin B was significantly enhanced after exposure to ozone.Conclusion:Although ozone was highly effective on the yeast form of C. albicans and it can inhibit the formation of germ tubes in C. albicans, the complete removal of biofilms did not happen even after 60 min. It seems that ozone therapy induces resistance to amphotericin B.

The efficacy of gaseous ozone against different forms of Candida albicans.

Ozone is an inorganic molecule with effective antimicrobial properties. Clinical treatment of ozonated water was used for the elimination of Candidaalbicans, Enterococcus faecalis, endotoxins, and biofilms from root canals. In addition, its therapeutic effects for tinea pedis, ulcers, and leishmaniasis were investigated. The purpose of the present study was to evaluate the fungicidal effects of ozone on different forms of C. albicans. In addition, antifungal susceptibility profile of strains was assessed before and after exposure to ozone. Fifty strains of C. albicans were exposed to gaseous ozone at different times. Furthermore, biofilm formation and germ tube production were evaluated when yeast suspensions were exposed to ozone. In addition, antifungal susceptibility of ozone resistant colonies was investiagted as compared to controls. Ozone was highly effective in killing C.albicans in yeast form and inhibition of germ tube formation during 210 and 180 s, respectively. Although with increasing exposure time biofilm production was considerably decreased, resistance to ozone was much higher among vaginal and nail isolates even after 60 min. All the strains were sensitive to fluconazole, caspofungin, and terbinafine pre- and post-ozone exposure. Resistance to amphotericin B was significantly enhanced after exposure to ozone. Although ozone was highly effective on the yeast form of C.albicans and it can inhibit the formation of germ tubes in C.albicans, the complete removal of biofilms did not happen even after 60 min. It seems that ozone therapy induces resistance to amphotericin B.

The effect of subinhibitory concentrations of gentian violet on the germ tube formation by Candida albicans and its adherence to oral epithelial cells

ObjectiveThis study investigated the effect of subinhibitory concentrations of gentian violet on the germ tube formation by Candida albicans and its adherence ability to oral epithelial cells. MethodsThirty strains of C. albicans isolated from denture wearers, normal healthy individuals and HIV positive patients were used in the study. The antifungal property (Minimum Fungicidal Concentration) of gentian violet was determined at various time intervals using a microdilution technique. The effect of subinhibitory concentrations of gentian violet on the adherence ability (0.000244%) and on germ tube formation ((0.000244%, 0.000122%, 0.000061% and 0.000031%) was determined. In both experiments, water was used as a control. The test results were compared using the Kruskal-Wallis test. ResultsAt 60min a high concentration (0.0078%) of gentian violet was required to completely kill C. albicans. Subinhibitory concentrations of gentian violet significantly reduced the adherence ability of C. albicans by 57% (p<0.01) and equally inhibited germ tube formation (p<0.01) compared with the controls. The inhibition was concentration dependent, with up to 98% reduction at a concentration of 0.000244%. Germ tube reduction was significantly higher in the isolates from the HIV positive patients than in the isolates from denture wearers. ConclusionAt high concentrations, gentian violet killed C. albicans, whereas at subinhibitory concentrations it reduced its virulence by preventing the adherence ability and germ tube formation. This suggests that the beneficial effects of gentian violet would last beyond the fungicidal concentrations in the treatment of candidiasis.

English

Medicinal plants are attractive sources in the search of bioactive compounds in the treatment of infectious diseases. Considering that the infectious agent often develops resistance to existing treatments rapidly, the searches for such compounds are a never-ending process. One such infection, caused by Candida spp. is candidiasis, which is a public health problem. Additionally, many strains have resistance to traditional therapies. Therefore, we tested the antifungal activity of four compounds present in Mentha spp. with promising antifungal precedents. We measured inhibition of growth by microdilution, disruption of biofilm viewed by electron microscopy, inhibition of germ tube formation by optical microscopy and toxicity on HaCaT cells. Tests showed that the compounds tested had antifungal activity with a minimum inhibitory concentration of 0.5 mg/mL, at least, 50% of biofilm inhibition in the 0.5 mg/mL concentration, an inhibition of polymorphism to 86% and the changes in the cell envelope of yeast (SEM) and cell viability above 50% among the Candida strains tested. Therefore, the compounds exhibit promising antifungal properties and provide a reasonable therapeutic window to be used in association with other traditional antimitotic. Key words: Carvone, menthone, menthofuran, pulegone, Candida spp., biofilm, cytotoxicity.