Anticandidal Compounds Research Articles

Exploring the Antifungal Activity and Action of Saussurea costus Root Extracts against Candida albicans and Non-albicans Species.

The isolation and assessment of the active constituents in polar and non-polar crude extracts of Saussurea costus roots as antifungal agents, against Candida albicans and non-C. albicans (NAC) species, was the aim of this current investigation. The SEM “Scanning electron microscopy” imaging provided potential action modes of n-hexane extract (nhhE) toward Candida spp., whereas the TLC-DB “Thin layer chromatography-direct bioautography” was employed for detecting the anticandidal compounds. nhhE had the greatest biocidal activity against all strains and clinical isolates of Candida spp. with maximum zones of inhibition. SEM revealed the occurrence of irregular, dense inclusions of C. albicans cell walls after treatment with nhhE for 12 h. Complete morphological distortions with lysed membranes and deterioration signs appeared in most treated cells of C. parapsilosis. The most effectual compound with anticandidal activity was isolated using TLC-BD and identified as sesquiterpene by GC/MS analysis. The infra-red analysis revealed the presence of lactone ring stretching vibrations at 1766.72 cm−1. The anticandidal activity of nhhE of S. costus roots was confirmed from the results, and the treated cotton fabrics with nhhE of S. costus possessed observable activity against C. albicans. Data could recommend the practical usage of S. costus extracts, particularly nhhE, as influential natural bioactive sources for combating pathogenic Candida spp.

The cytotoxicity studies of phytosterol discovered from Rhizophora apiculata against three human cancer cell lines

Gallic acid: An anticandidal compound in hydrolysable tannin extracted from the barks of Rhizophora apiculata BlumeLim Sheh Hong, Darah Ibrahim, Jain Kassim, Suraya Sulaiman

Preparation and characterization of gallic acid-loaded PLGA hydrogel as teat sealant for preventing mastitis in dry cows

Gallic acid: An anticandidal compound in hydrolysable tannin extracted from the barks of Rhizophora apiculata BlumeLim Sheh Hong, Darah Ibrahim, Jain Kassim, Suraya Sulaiman

ENDOPHYTIC BACTERIA DIVERSITY FROM ZINGIBERACEAE AND ANTICANDIDAL CHARACTERIZATION PRODUCED BY Pseudomonas helmanticensis

Endophytic microbes are sources for the novel antibiotic. We isolated endophytic bacteria from Zingiberaceae collected from West Sulawesi, Indonesia, and investigated their anticandidal activity. Molecular identification of the isolates was done using 16S rRNA gene sequence analysis. The antimicrobial activity was tested against four bacteria and one yeast. The anticandidal compound of selected bacteria was extracted using three different solvents (chloroform, ethyl acetate, and methanol), and each fraction was tested for their anticandidal activity. Anticandidal minimum inhibitory concentration (MIC) was determined with concentration ranging from 300 to 18.75 μg/mL, and the morphology of the Candida cells after treatment was confirmed by scanning electron microscope (SEM). The identification of anticandidal compounds was conducted using GC-MS. A total of 24 isolates were collected from Zingiberaceae plants. There were 14 genera and 19 species belonging to Gammaproteobacteria (66.67%), Alphaproteobacteria (25.00%), Actinobacteria (4.17%), Bacteriodetes (4.17%), and a new record for Lelliottia aquatilis as an endophytic bacteria. One of 24 isolates identified as Pseudomonas helmanticensis isolated from Alpinia melichroa showed anticandidal activity. Ethyl acetate was the appropriate solvent to extract the anticandidal compounds. Diisooctyl phthalate was found as the most abundant compound in the extract for the anticandidal activity. An increase in extract concentration did not reduce the Candida cell number. The extract treatment showed membrane disruption of Candida albicans cells. We propose that active compounds from P. helmanticensis are potential as anticandidal sources and could be explored more for the pharmaceutical industry.

In Vitro and In Vivo Anti-Candida spp. Activity of Plant-Derived Products.

Candidiasis therapy, especially for candidiasis caused by Candida non-albicans species, is limited by the relatively reduced number of antifungal drugs and the emergence of antifungal tolerance. This study evaluates the anticandidal activity of 41 plant-derived products against Candida species, in both planktonic and biofilm cells. This study also evaluates the toxicity and the therapeutic action of the most active compounds by using the Caenorhabditis elegans–Candida model. The planktonic cells were cultured with various concentrations of the tested agents. The Cupressus sempervirens, Citrus limon, and Litsea cubeba essential oils as well as gallic acid were the most active anticandidal compounds. Candida cell re-growth after treatment with these agents for 48 h demonstrated that the L. cubeba essential oil and gallic acid displayed fungistatic activity, whereas the C. limon and C. sempervirens essential oils exhibited fungicidal activity. The C. sempervirens essential oil was not toxic and increased the survival of C. elegans worms infected with C. glabrata or C. orthopsilosis. All the plant-derived products assayed at 250 µg/mL affected C. krusei biofilms. The tested plant-derived products proved to be potential therapeutic agents against Candida, especially Candida non-albicans species, and should be considered when developing new anticandidal agents.

Lipidomic Insight of Anticandidal Perillyl Alcohol and Sesamol Induced Candida Membrane Disruption: Implications of Lipid Alteration, Impaired Fluidity and Flippase Activity.

Considering the emergence of multidrug resistance (MDR) in prevalent human fungal pathogen, Candida albicans, there is parallel spurt in the development of novel strategies aimed to disrupt MDR. The cell envelope of C. albicans comprises a wealth of lipid moieties contributing towards long-term survival of pathogen that could be exploited as efficient antifungal target owing to the advancements made in mass spectrometry based lipidomics technology. This study aimed to utilize the lipidomics approach to unveil several lipid-associated changes in response to two natural anticandidal compounds perillyl alcohol (PA) and sesamol (Ses). Lipidomics is performed through ESI-MS, flippase activity by FACS, fluorescence spectrometric analysis is used to assess membrane fluidity. Lipidomic analyses revealed that phosphatidylcholine (PtdCho) were decreased in the presence of Ses with considerable differences at specie level. Concurrently, we explored increased inward translocation (flip) of fluorophore labelled PtdCho across the plasma membrane attributed to enhanced PtdCho specific flippase activity. A considerable decrement in phosphatidylethanolamine (PtdEtn) leading to altered membrane fluidity was observed in response to PA and Ses. Additionally, we could detect alteration in the levels of phohatidylserine (PtdSer) and phosphatidylglycerol (PtdGro) along with decreased triacylglycerides (TAG). The differential expressions of various lipid biosynthetic pathway genes by RT-PCR corroborated with the lipidomics data. Furthermore, PA and Ses leads to potentiation of membrane targeting drugs (azole and polyene) and displayed additive effect. Our work offers the basis of further understanding the regulation of lipid homeostasis in C. abicans so that better therapeutic targets could be identified to combat MDR.

Candida albicans Infection Model in Zebrafish (Danio rerio) for Screening Anticandidal Drugs.

Candida albicans is an opportunistic fungal pathogen which causes systemic infections in human. In this study, C. albicans infection model was developed in zebrafish to understand the host-pathogen interactions for straightforward anticandidal drug screening. To develop the infection, 1 × 106cells of C. albicans suspended in phosphate-buffered saline were deposited in zebrafish dorsal muscle by manually operated syringe. The infection progression was externally assessed by a scale of wound-healing events, based on visible changes of yeast deposited in the muscle tissues. Chemotherapy was carried out with known antifungal drugs (fluconazole, nystatin, and amphotericin B) and a potential antifungal agent, chitosan silver nanocomposites (CAgNC), after the infection as direct exposure in the water. Histopathological analysis was performed to identify the pathogen virulence and the host-pathogen interaction during the infection. The light microscopic observations and histopathological analysis revealed the yeast-hyphae transition at the site of infection (at 72hpi) and progression of the infection in the host tissues. The larval survival rate under fluconazole (up to 80μgmL-1) and nystatin (up to 20μgmL-1) was > 90% and for CAgNC it was 40% at 36h post-exposure (hpe). The infection progression was suppressed with the fungicidal treatments. Among inflammatory genes, il-1β has been highly upregulated (14.68-fold) at 24h post infection (hpi). Both il-1β and tnf-α were moderately upregulated in infected fish gills at 72hpi. Among the C. albicans antioxidant genes, cat1 and sod2 have been upregulated during the infection, and relative expression folds were increased from low to moderate levels with the time. We demonstrate the approach for the development of artificial infection model of zebrafish with C. albicans. By this mini vertebrate zebrafish model, researchers will be able to study novel anticandidal compounds in vivo with respect to the host, pathogen, and their interactions.

Anticandidal Potential of Stem Bark Extract from Schima superba and the Identification of Its Major Anticandidal Compound.

Plant-derived extracts are a promising source of new drugs. Schima superba is traditionally used in China for heat clearing, detoxification, and treatment of furuncles. In this study, the anticandidal properties and mechanism of action of S. superba (SSE) were explored using a stem bark extract. SSE possessed high polyphenol and saponin contents of 256.6 ± 5.1 and 357.8 ± 31.5 µg/mg, respectively. A clear inhibition zone was observed for C. albicans growth through the disc diffusion method and the 50% inhibition of C. albicans by SSE was 415.2 µg/mL. Transcriptomic analysis in C. albicans treated with different doses of SSE was conducted through RNA-seq. Average values of 6068 genes and 20,842,500 clean reads were identified from each sample. Among these samples, 1680 and 1956 genes were differentially expressed genes (DEGs) from the SSE treatments of 0.2 and 0.4 mg/mL, respectively. C. albicans growth was inhibited by the changes in gene expression associated with the cell wall and membrane composition including the regulation of chitin degradation and ergosterol biosynthesis. This result could be reflected in the irregularly wrinkled morphology of the ruptured cell as revealed through SEM analysis. ESI-MS and NMR analyses revealed that the major compound purified from SSE was sasanquasaponin III and the 50% inhibition of C. albicans was 93.1 µg/mL. In summary, the traditional Chinese medicine S. superba can be applied as an anticandidal agent in complementary and alternative medicine.

Anti-candidal activity of Aspergillus flavus IBRL-C8, an endophytic fungus isolated from Cassia siamea Lamk leaf

Gallic acid: An anticandidal compound in hydrolysable tannin extracted from the barks of Rhizophora apiculata BlumeLim Sheh Hong, Darah Ibrahim, Jain Kassim, Suraya Sulaiman

Endophytic Diaporthe sp. ED2 Produces a Novel Anti-Candidal Ketone Derivative.

This study aimed to examine the anti-candidal efficacy of a novel ketone derivative isolated from Diaporthe sp. ED2, an endophytic fungus residing in medicinal herb Orthosiphon stamieus Benth. The ethyl acetate extract of the fungal culture was separated by open column and reverse phase high-performance liquid chromatography (HPLC). The eluent at retention time 5.64 min in the HPLC system was the only compound that exhibited anti-candidal activity on Kirby-Bauer assay. The structure of the compound was also elucidated by nuclear magnetic resonance and spectroscopy techniques. The purified anti-candidal compound was obtainedas a colorless solid and characterized as 3-hydroxy-5-methoxyhex-5-ene-2,4-dione. On broth microdilution assay, the compound also exhibited fungicidal activity on a clinical strain of Candida albicans at a minimal inhibitory concentration of 3.1 μg/ml. The killing kinetic analysis also revealed that the compound was fungicidal against C. albicans in a concentration- and time-dependent manner. The compound was heat-stable up to 70°C, but its anti-candidal activity was affected at pH 2.

A Novel Anticandidal Compound Containing Sulfur from Endophytic Fungus Emericella sp.

Background: Endophtes live symbiotically in the tissues of host plants without causing visible harm. These endophytes produce not only the plant bioactive metabolites but also a host of unexplored structurally diverse natural products having biological activity. New niches may be explored for bioprospecting endophytes for the development of novel candidate molecules having industrial applications. Methods: The bioactive compound is isolated from the endophyte Emericella sp. associated with Azadirachta indica A. Juss (Neem), that potentially inhibits or prevents the human pathogenic disease caused by Candida albicans. The molecule was characterized by employing TLC, IR and GCMS. The isolated compound was identified as N-[(2S, 3aR, 6S, 7aS)-6-acetamido-octahydro-1,3-benzothiazol-2-yl]2- (adamantan-1-yl) acetamide having sulfur. Results: In the present study, a new molecule containing sulphur inhibits the growth of Candida specifically. The molecule can serve as a lead molecule for designing new drugs having anticandidal activity. The compound having anticandidal activity belongs to the class of antifungal agent – allylamine, azole and polyene. Conclusion: A new molecule containing sulphur and inhibiting specifically C. albicans is reported in the present study. They are known to promote plant growth and yield, act as biocontrol agents and are known to produce secondary metabolites, such as enzymes, growth hormones, antimicrobial, antifungal, or anticancerous substances. Keywords: Anticandidal drug, Azadirachta indica, Candida albicans, Emericella, endophyte.

Marine sources hold treasure for therapeutics

Tuberculosis and Candiasis are the two infectious diseases which globally affects considerable number of the population annually. Many synthetic drugs are accessible for the treatment but are expensive and cause toxicity in humans. Moreover, prolong exposure of the drugs has led to emergence of multidrug resistance. Marine derived natural compounds as alternative strategies have attracted significant attention and been widely used nowadays. Pharmacologic researches with marine compounds have contributed potentially to the ongoing global search for therapeutic agents in the treatment of multiple diseases. We present an overview of some promising marine bioactive antimycobacterial and anticandidal compounds from a therapeutic point of view isolated from marine sources.

Screening for Anticandidal and Antibiofilm Activity of Some Herbs in Thailand

Purpose: To evaluate the anticandidal activity of the ethanol extracts of 12 herbs from Thailand. Methods: The herbs studied were Alpinia galanga, Curcuma longa, Curcuma zedoaria, Mentha cordifolia, Ocimum africanum, Ocimum basilicum, Ocimum sanctum, Piper betle, Piper chaba, Piper nigrum, Piper sarmentosum and Zingiber officinale. Various Candida spp. were examined for minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) using microdilution method; time-kill assay was also used to assess the plants. Antibiofilm activity was investigated using a 3-[4, 5dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide (MTT assay). Gas chromatography mass spectrometry (GC-MS) analysis, thin layer chromatography (TLC) fingerprinting and TLC-bioautography were used to determine the active anticandidal compounds. Results: All tested herbs, except extracts of P. nigrum and Limiaceae family, showed varying zones of inhibition against Candida albicans ATCC 90028. P. betle revealed the strongest anticandidal activity against all tested strains with MIC ranging from 1.56 to 3.13 mg/ml, and MFC from 3.13 to 8.33 mg/ml. Killing activity depended on time and concentrations of the extract. The concentration of P. betle extract required to inhibit ≥ 90 % biofilm formation of C. albicans ATCC 90028 was 3.13 ± 0.15 mg/ml, while that to remove ≥ 90 % biofilm growth was 12.50 ± 0.69 mg/ml. The result of GC-MS analysis showed the major compound of P. betle extract responsible for anticandidal activity as 4-chromanol. Conclusion: P. betle extract contains 4-chromanol which is a good potential anticandidal agent for the treatment of oral infectious diseases caused by certain Candida spp.

Isolation and identification of anticandidal compound from Streptomyces sp. VITPK9

Candida infections are frequently reported in both HIV and cancer patients. Recent reports have shown that Candida participates in malignant transformation of oral fibrosis. The aim of the present study was to isolate and to identify anticandidal compound from soil Streptomyces sp. VITPK9. It was isolated from a brine spring of Manipur located in Thoubal district, Manipur, India. The ethyl acetate extract from culture supernatant of Streptomyces sp. VITPK9 was prepared and purified by silica gel column chromatography and HPLC. The purified compound was identified by using 1H and 13C NMR spectral data and based on the similarity index with reference compounds available in the mass spectra library of National Institute for Standards and Technology as pyrrolo[1,2-a]pyrazine-1,4-dione,hexahydro-3-(phenylmethyl)-. The antifungal activity of the purified compound was tested against the Candida strains according to the National Committee for Clinical Laboratory Standards guidelines and it was revealed that its MIC50 value ranged from 0.78 to 2.00 μg/mL. The results of the study suggest that Streptomyces sp. VITPK9 is the potential source for diketopiperazine type of anticandidal compounds.

Synthesis and biological evaluation of some hydrazone derivatives as new anticandidal and anticancer agents

New hydrazone derivatives were synthesized via the nucleophilic addition–elimination reaction of 2-[(1-methyl-1H-tetrazol-5-yl)thio)]acetohydrazide with aromatic aldehydes/ketones. The compounds were tested in vitro against various Candida species and compared with ketoconazole. Genotoxicity of the most effective anticandidal compounds was evaluated by umuC and Ames assays. All compounds were also investigated for their cytotoxic effects on NIH3T3 and A549 cell lines. Compound 8 was the most effective antifungal derivative against C. albicans (ATCC-90028) with a MIC value of 0.05 mg/mL. Compound 5 can be identified as the most promising anticancer agent against A549 cancer cell lines due to its inhibitory effect on A549 cell lines and low toxicity to NIH3T3 cells.

Enhancement of anti-candidal activity of endophytic fungus Phomopsis sp. ED2, isolated from Orthosiphon stamineus Benth, by incorporation of host plant extract in culture medium

This study examined the effect of host extract in the culture medium on anti-candidal activity of Phomopsis sp. ED2, previously isolated from the medicinal herb Orthosiphon stamineus Benth. Interestingly, upon addition of aqueous host extract to the culture medium, the ethyl acetate extract prepared from fermentative broth exhibited moderate anti-candidal activity in a disc diffusion assay. The minimal inhibitory concentration of this extract was 62.5 μg/ml and it only exhibited fungistatic activity against C. albicans. In the time-kill study, a 50% growth reduction of C. albicans was observed at 31.4 h for extract from the culture incorporating host extract. In the bioautography assay, only one single spot (Rf 0.59) developed from the extract exhibited anti-candidal activity. A spot with the a similar Rf was not detected for the crude extract from YES broth without host extract. This indicated that the terpenoid anti-candidal compound was only produced when the host extract was introduced into the medium. The study concluded that the incorporation of aqueous extract of the host plant into the culture medium significantly enhanced the anti-candidal activity of Phomopsis sp. ED2.

Anticandidal activity of Cassia fistula and its effect on ergosterol biosynthesis

Context: Cassia fistula Linn. (Caesalpiniaceae) has been used in folk medicine. Anthraquinone derivative rhein having antimicrobial properties is actively present in C. fistula fruit. Although, as yet there has been no study of its anticandidal potential.Objective: The present study was conducted to determine the phytochemical composition of fruit pulp and seed extract and their effect on Candida albicans ATCC 10261, Candida glabrata ATCC 90030 and Candida tropicalis ATCC 750, respectively.Materials and methods: The fruit pulp and seed extracts were tested for phytochemicals by various standard methods and rhein was identified by thin-layer chromatography. The anticandidal activity was determined by minimum inhibitory concentration (MIC), growth curve studies, cytotoxicity and ergosterol estimation assay.Results: The fruit pulp and seed extracts showed high content of phenolic compounds. Rhein was identified in both extracts, Rf 0.38. MICs of seed extract obtained with C. albicans, C. tropicalis and C. glabrata is 350, 300 and 300 μg/ml. However, for fruit pulp extract, these values significantly reduced to 150, 250 and 100 μg/ml, respectively. Comparative MIC values for fluconazole were 16, 16 and 04 µg/ml. At MICs, pulp reduced ergosterol content in cell membrane of C. albicans, C. tropicalis and C. glabrata by 54.42, 48.78 and 68.0%, seed extract by 38.11, 47.0 and 45.0%, whereas, fluconazole showed 93.56, 89.21 and 98.0%, respectively.Discussion and conclusion: C. fistula fruit pulp and seed extract possessed anticandidal activity. The result was significantly correlated between the MICs, cytotoxicity and ergosterol inhibition. It was concluded that the crude extract is a promising source for anticandidal compounds.

Frequency of Interleukin-4 (IL-4) -589 Gene Polymorphism and Vaginal Concentrations of IL-4, Nitric Oxide, and Mannose-Binding Lectin in Women with Recurrent Vulvovaginal Candidiasis

A C-->T substitution at position -589 in the interleukin-4 (IL-4) gene is associated with increased production of IL-4. Associations between this polymorphism and recurrent vulvovaginal candidiasis (RVVC), as well as vaginal concentrations of IL-4 and the anticandidal compounds nitric oxide (NO) and mannose binding lectin (MBL), were evaluated. Vaginal samples obtained by lavage from 42 women with RVVC during the acute stage of the disease and 43 control samples were assayed by enzyme-linked immunosorbent assay for IL-4 and NO metabolites. The -589 IL-4 gene polymorphism was detected by polymerase chain reaction and endonuclease digestion. Data were analyzed by Fisher's exact test, the nonparametric Mann-Whitney and Kruskal-Wallis tests, and Spearman rank correlation. P < .05 was considered significant. Candida albicans was identified in 38 patients with RVVC; 3 others had infection due to Candida tropicalis, and 1 had infection due to Candida krusei. The IL-4 T,T genotype was detected in 59.5% of patients with RVVC and in 7.0% of control subjects (P < .0001). The frequency of IL-4*T was 76.2% in patients with RVVC and 23.3% in control subjects (P < .0001). The median concentration of vaginal IL-4 was elevated in patients with RVVC, compared with control subjects (P < .0001). Conversely, vaginal concentrations of NO metabolites (P = .02) and MBL (P < .0001) were reduced in patients with RVVC. There was a positive association between IL-4*T homozygosity and vaginal IL-4 levels (P < .0001) and negative associations between this genotype and vaginal NO (P = .01) and MBL (P < .0001) concentrations. Reduced vaginal levels of anticandidal factors in IL-4*T homozygotes may increase susceptibility to RVVC.

Cell cycle effects of the phenothiazines: trifluoperazine and chlorpromazine in Candida albicans

The study demonstrates the in vitro effectiveness of phenothiazine compounds, i.e. chlorpromazine and trifluoperazine against Candida albicans. Anticandidal effect of these drugs is suggested to be because of their interaction with Ca 2+/calmodulin dependent protein phosphorylation. 3H-thymidine uptake studies revealed that both these compounds affect the DNA synthesis along with decrease in activities of nuclear calmodulin (CaM) and Ca 2+/calmodulin dependent protein kinase (CaMPK). Failure in cell growth was due to defect in CaM mediated cell cycle arrest. Flow cytometric analysis showed that progression through G 1 and mitotic phase was affected when cells after α-factor arrest were grown in the presence of chlorpromazine or trifluoperazine. These drugs also produced significant decline in the cellular lipids and phospholipids. 14C-acetate incorporation studies further substantiated these results. We suggest that chlorpromazine or trifluoperazine affect the cell cycle through DNA synthesis ( S phase) and cell division phases which are governed by calmodulin and Ca 2+/calmodulin dependent protein phosphorylation and lipids and phospholipids appear to be additional targets of phenothiazine compounds in C. albicans. These results will have important significance in the development of new anticandidal compounds.

Cell cycle effects of the phenothiazines: trifluoperazine and chlorpromazine in Candida albicans.

The study demonstrates the in vitro effectiveness of phenothiazine compounds, i.e. chlorpromazine and trifluoperazine against Candida albicans. Anticandidal effect of these drugs is suggested to be because of their interaction with Ca(2+)/calmodulin dependent protein phosphorylation. 3H-thymidine uptake studies revealed that both these compounds affect the DNA synthesis along with decrease in activities of nuclear calmodulin (CaM) and Ca(2+)/calmodulin dependent protein kinase (CaMPK). Failure in cell growth was due to defect in CaM mediated cell cycle arrest. Flow cytometric analysis showed that progression through G(1) and mitotic phase was affected when cells after alpha-factor arrest were grown in the presence of chlorpromazine or trifluoperazine. These drugs also produced significant decline in the cellular lipids and phospholipids. 14C-acetate incorporation studies further substantiated these results. We suggest that chlorpromazine or trifluoperazine affect the cell cycle through DNA synthesis (S phase) and cell division phases which are governed by calmodulin and Ca(2+)/calmodulin dependent protein phosphorylation and lipids and phospholipids appear to be additional targets of phenothiazine compounds in C. albicans. These results will have important significance in the development of new anticandidal compounds.