Synergistic potential of Bauhinia holophylla leaf extracts with conventional antifungals in the inhibition of Candida albicans: A new approach for the treatment of oral candidiasis

Background: Green Tea is made from the leaf of the plant “Camellia sinensis”. Green tea is reported to contain thousands of bioactive ingredients including catechins which have shown great promise for having antimicrobial effects. Periodontal diseases represent one of the most prevalent diseases around the world and the main etiologic factor behind it, is plaque accumulation, in addition certain kinds of bacteria have been detected frequently in subjects suffering from periodontitis, Several studies suggested that the outcome of periodontal treatment is better if particular pathogens including Aggregatibacteractinomycetemcomitans can no longer be detected after therapy. Materials and Methods: plaque samples were collected from 20 patients suffering from chronic periodontitis with probing pocket depth of at least 6 mm, Aggregatibacteractinomycetemcomitans (A.A) was isolated and diagnosed according to morphological characteristics and biochemical tests. Green tea leaves were extracted by using water and alcohol. The first experiment involved testing the sensitivity of A.A to different concentrations of the extracts using agar well diffusion method,the second experiment involved determination of the minimum inhibitory concentration and then determination of the minimum bactericidal concentration of the extract against the bacteria, laboratory analysis of green tea extracts using high pressure liquid chromatography (HPLC) was performed. Results: Both green tea extracts were effective in inhibition of Aggregatibacteractinomycetemcomitans using agar well diffusion method, 90% and 100% concentrations of alcoholic extract showed larger inhibition zones than chlorhexidinegluconate 0.2% with statistically significant difference, CHX showed higher inhibition zones than all aqueous extract concentrations.The MIC (minimum inhibitory concentration) of alcoholic green tea extract that inhibit Aggregatibacteractinomycetemcomitans growth was 60%, The MIC of aqueous green tea extract that inhibits Aggregatibacteractinomycetemcomitans growth was 70%.The MBC (minimum bactericidal concentration) of alcoholic green tea extract that killsAggregatibacteractinomycetemcomitans was 80%, the MBC of aqueous green tea extract that kills Aggregatibacteractinomycetemcomitans growth was 90%. HPLC analysis of aqueous and alcoholic green tea extracts revealed that alcoholic extract contained higher concentration of EGCG while aqueous extract had higher content of catechin and epicatechin. Conclusion: Green tea extracts were effective against Aggregatibacteractinomycetemcomitans, alcoholic green tea extract showed inhibition ability more than the aqueous green tea extract and more than CHX and it showed bactericidal activity at 80%,90% and 100% concentrations.

β-Defensins (BDs) are highly conserved antimicrobial peptides important in innate defence against bacteria. β-Defensin 3 has a specific role in protecting the skin. This study quantified the minimal inhibitory concentration (MIC) of human (h)BD3 against Staphylococcus pseudintermedius isolates from atopic and healthy dogs. Single colony isolates (1 × 10(5) colony-forming units/mL log phase) were cultured with doubling dilutions of hBD3 in sodium phosphate buffer from 0.8 to 50 μg/mL at 37 °C for 2 h, before adding 100 μL of tryptone soy broth and incubating for a further 20 h. Bacterial growth was assessed as the mean optical density at 540 nm corrected for background. The median MIC was 12.5 μg hBD3/mL (range 3.125-25 μg/mL; n=22). Forty-five percent of the isolates were inhibited at ≤ 6.25 μg hBD3/mL, and 90% were inhibited at ≤ 12.5 μg hBD3/mL. Bacterial growth was not inhibited at ≤ 1.6 μg hBD3/mL. There were no significant differences in the inhibition by hBD3 of isolates from atopic (median MIC 12.5 μg/mL, range 6.25-25 μg/mL, n=14) and healthy dogs (median MIC 9.4 μg/mL, range 3.125-12.5 μg/mL, n = 8); from noninfected colonized sites (median MIC 12.5 μg/mL, range 3.125-25 μg/mL, n=16) and infected lesions (median MIC 9.4 μg/mL, range 6.25-12.5 μg/mL, n=6); or between sample sites (nose median MIC 12.5 μg/mL, range 6.25-25 μg/mL, n=5; perineum median MIC 12.5 μg/mL, range 3.125-25 μg/mL, n=7; ear median MIC 6.25 μg/mL, range 6.25-12.5 μg/mL, n=4; lesions median MIC 9.4 μg/mL, range 6.25-12.5 μg/mL, n=6). In conclusion, hBD3 inhibited the growth of canine S. pseudintermedius isolates in vitro irrespective of origin.

To evaluate the cytotoxicity of crude aqueous and ethanol extracts of Zaleya pentandra against oestrogen receptor-positive breast cancer cell line Michigan Cancer Foundation-7. The study was conducted at the Institute of Chinese Medicine, the Chinese University of Hong Kong, Hong Kong, from March to September 2017, and comprised Zaleya pentandra herbaceous perennial plant collected from Pakistani cities of Shakargarh, Lahore and Sialkot. Both aqueous and ethanol extracts were prepared in solvents following Soxhlet extraction technique. Rate of reduction in viability of cancer cell line was studied through MTT (dye compound 3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assay on the basis of time of incubation and the dose of extract. Analysis was performed using High Performance Liquid Chromatography with Diode Array Detector in order to found the compounds responsible for the death of cancer cells. Cell viability was observed to be dose-dependent (range: 16.7-37.4%) for aqueous extracts. Minimum inhibition concentration was 16.65% at 200μg/ml after 24 hours of incubation, whereas maximum inhibition was 37.39% at 3200μg/ml. Ethanol extracts showed less inhibition, with maximum inhibition being 25.29% at 1000μg/ml and minimum 13.57% at 62.5μg/ml. Certain polar compounds, like Hydroxytrosol and Tyrosol, could be obtained from the aqueous extracts only. Zaleya pentandra aqueous extract was found to have potential benefit towards cytotoxicity of breast cancer cells.

Objective: To chemically characterize Spondias mombin L. extracts and to evaluate the in vitro antimicrobial activity of these extracts on Enterococcus faecalis and Pseudomonas aeruginosa. Material and methods: Both extracts were submitted to phytochemical prospecting and thin layer chromatography (TLC) analysis prior to determination of Minimum Inhibitory Concentration (MIC), Minimum Inhibitory Adhesion Concentration (MIAC) and Bactericidal Kinetics (BK). The Student’s t-test was used to compare MIC results at the 5% significance level. The chemical characterization, MIAC and BK data were analyzed descriptively. Results: Phenolic compounds, flavonoids, tannins and saponins were detected in the extracts. The hydroethanolic extracts in different concentrations showed statistically superior inhibitory bacterial growth activity (p <0.05) compared to the control against the two investigated microorganisms, whereas the aqueous extracts (also in different concentrations) was statistically superior (p <0.05) to the control only against Pseudomonas aeruginosa. The anti-adherent effect of both extracts was superior to that of the control for P. aeruginosa, and similar to that for E. faecalis. In their crude and diluted concentrations, aqueous and hydroethanolic extracts showed bactericidal action within the first two to four hours of contact with the bacteria. Conclusion: Spondias mombin L. extracts in different concentrations showed superior antimicrobial activity than 0.12% chlorhexidine digluconate against the microorganisms with this activity associated to the chemical characterization of these extracts.

Chlorhexidine is a highly effective topical broad-spectrum agent against Candida spp.

The in vitro antilisterial activities and time kill regimes of crude aqueous and n-hexane extracts of the husk fiber of Cocos nucifera were assessed. The aqueous extracts were active against 29 of the 37 test Listeria isolates while the n-hexane extracts were active against 30. The minimum inhibitory concentrations (MICs) of all the susceptible bacteria ranged between 0.6 and 2.5 mg/ml for the aqueous fraction and between 0.6 and 5.0 mg/ml for the n-hexane extract. The average log reduction in viable cell count in the time kill assay ranged between 0.32 Log10 and 3.2 Log10cfu/ml after 4 h of interaction, and between 2.6 Log10 and 4.8 Log10 cfu/ml after 8 h interaction in 1 × MIC and 2 × MIC (aqueous extract); and between 2.8 Log10 and 4.8 Log10 cfu/ml after 4 h of interaction, and 3.5 Log10 to 6.2 Log10 cfu/ml after 8 h interaction in 1 × MIC and 2 × MIC for the n-hexane extract. The extract was bactericidal against one of the test bacteria at 1 × MIC and against three of the test bacteria at 2 × MIC for the 8 h interaction period for the aqueous extract, while for the n-Hexane fraction; the extract was bactericidal against all the five test bacteria at both MICs after the 8 h interaction period. We suggested that the crude aqueous and n-hexane extracts of the husk of C. nucifera could be bacteriostatic or bactericidal depending on the time of exposure and concentration. Key words: Cocos nucifera, n-hexane extract, aqueous extract, minimum inhibitory concentration, time-kill.

ABSTRACTFicus deltoidea Jack (Moraceae) is a well-known medicinal plant used in customary medication among the Malay people to reduce and mend sicknesses such as ulcers, psoriasis, cytotoxicity, cardioprotective, inflammation, jaundice, vitiligo, hemorrhage, diabetes, convulsion, hepatitis, dysentery injuries, wounds, and stiffness. Ficus deltoidea contains a wide variety of bioactive compounds from different phytochemical groups such as alkaloids, phenols, flavonoids, saponins, sterols, terpenes, carbohydrates, and proteins. The genus Ficus has several hundreds of species, which shows excellent therapeutic effects and a wide variety of helpful properties for human welfare. Searching information was collected by using electronic databases including Web of Science, Science Direct, Springer, SciFinder, PubMed, Scopus, Medline, Embase, and Google Scholar. This review is, therefore, an effort to give a detailed survey of the literature on its pharmacognosy, phytochemistry, phytochemical, and pharmacological properties of Ficus and its important species. This summary could be beneficial for future research aiming to exploit the therapeutic potential of Ficus and its useful medicinal species.

Aeromonas spp. are gram-negative bacilli of the Gammaproteobacteria class and Aeromonadaceae family. These bacteria are a public health challenge, particularly in South America. They harbor chromosomal β-lactamases belonging to classes B, C, and D, whose presence varies among species. This retrospective study aimed to evaluate the minimum inhibitory concentrations (MICs) of piperacillin-tazobactam (PTZ) and cefotaxime (CTX) for clinical strains of Aeromonas spp. A secondary objective was to assess the resistance profile of these strains to cefepime (FEP), aztreonam (AZT), ciprofloxacin (CIP), levofloxacin (LEV), and trimethoprim-sulfamethoxazole (SXT). The study was conducted at the Microbiology Laboratory of Cayenne Hospital (French Guiana) from January 2020 to December 2023. All isolated clinical Aeromonas strains, regardless of species or sample type, were included. In total, 99 strains were analyzed, comprising Aeromonas hydrophila (n = 79), Aeromonas caviae (n = 10), Aeromonas veronii (n = 6), and other Aeromonas spp. (n = 4). The median PTZ MIC was 0.5 mg/L for Aeromonas spp. overall, with six strains (6%) exhibiting MICs of >8 mg/L. For A. hydrophila, the median PTZ MIC was also 0.5 mg/L, lower than the median reported by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) for strains of 4 mg/L. For CTX, the median MIC for all Aeromonas spp. was 0.064 mg/L, with four strains (4%) having MICs of >4 mg/L. The median ceftazidime (CAZ) MIC was 0.125 mg/L. No resistance to AZT was detected, and 3% high-dose susceptibility to FEP was found. Resistance rates to other antibiotics were low: 6% to CIP, 6% to LEV, and 6% to SXT. In conclusion, our study demonstrated low resistance rates in Aeromonas spp. to PTZ (6%), CTX (4%), and CAZ (4%). These findings suggest that PTZ and CTX could be effective against Aeromonas infections, despite the lack of established clinical breakpoints by EUCAST.IMPORTANCEAeromonas spp. are clinically significant, particularly in tropical climates. This study provides essential MIC distribution data for piperacillin-tazobactam and cefotaxime, offering valuable insights for guiding empirical treatment choices. Understanding these susceptibility patterns is crucial for optimizing antimicrobial therapy, particularly in regions with limited resistance surveillance and standardized guidelines. By analyzing a large collection of clinical isolates, our findings contribute to evidence-based decision-making for Aeromonas infections and emphasize the importance of continued surveillance of antimicrobial susceptibility. These findings suggest that PTZ and CTX may be therapeutic options for Aeromonas infections, supporting their potential use in clinical practice.

Treatment failure in typhoid fever with ciprofloxacin susceptible Salmonella enterica Serotype Typhi

IntroductionBecause of resistance and side effects to common antifungal drugs activity, the research on herbal substances with antifungal activity is frequent. Lemon verbena (Lippia citriodora) is a member of Verbenaceae family. The aim of this study was to determine the anti-candida activities of the ethanolic and aqueous extracts of the lemon verbena leaves and compare them with nystatin and fluconazole.MethodsIn this 2015 study, 15 clinical isolates and standard strain of candida albicans PTCC 5027 were used, and the inhibitory effects of the ethanolic and aqueous extracts, Nystatin and Fluconazole, were evaluated using disk and well diffusion methods. Also, the minimal inhibitory concentration (MIC) was determined. Five concentrations of aqueous and ethanolic extracts (156–2500 μg/ml), Nystatin (8–128 μg/ml) and Fluconazole (4–64 μg/ml) were used in disk and well diffusion methods, and nine concentrations of aqueous and ethanolic extracts (19–5000 μg/ml), Nystatin (0.5–128 μg/ml), and Fluconazole (0.25–64 μg/ml) were applied for MIC. Data were analyzed using Tukey’s post-hoc and one-way ANOVA tests. The significant level was considered p < 0.05 in the current study.ResultsIn the well and disk diffusion techniques, limited growth inhibition halos were produced around some clinical isolates at different concentrations of ethanolic extract; however, no growth inhibitory halo was observed with any concentrations of the aqueous extract. The MIC values of ethanolic extract, aqueous extract, Nystatin and Fluconazole for clinical isolated and standard strain were 833 ± 78.5and 625μg/ml; 4156 ± 67.4 and 2500 μg/ml; 10.13 ± 1.91 and 4 μg/ml; and 1.97 ± 0.25 and 1 μg/ml, respectively.ConclusionThe results showed that the ethanolic extract was stronger than the aqueous extract of this plant, which can be used as an alternative for drugs. It is recommended that the ethanolic extract of this plant be investigated in vivo for better evaluation of its efficacy and properties.

BackgroundThe use of botanical drugs for treating various disorders has gained increasing attention in recent years, with many studies highlighting the efficacy of botanical antifungals against oral candidiasis. However, there is no definitive evidence indicating whether the botanical antifungals have superior or inferior efficacy compared to the conventional antifungals. This systematic review and meta-analysis evaluated the effectiveness of herbal and botanical antifungals versus conventional antifungals in treating oral candidiasis. This is the first pairwise comparison of the clinical efficacy.MethodsFrom inception to June 2024, PubMed, EMBASE, Scopus, and Web of Science were searched for randomized clinical trials published in English that investigated botanical antifungals compared to conventional antifungals in treating oral candidiasis. The primary outcome was lesion improvement, with in vitro Candida examination as the additional outcome. The lesion improvements were defined as the treatment duration (≤15 days and >15 days). Three independent reviewers screened the papers, and quality was assessed using Cochrane’s Risk of Bias two tools. For the Risk of Bias, five domains were evaluated and classified into three categories: low risk, some concerns, and high risk. A meta-analysis was conducted using STATA version 16 (Texas, United States). The protocol was registered in PROSPERO with an ID of CRD42024589391.ResultsFrom 1,595 studies identified, 10 trials were included with 426 patients, and 13 botanical drugs were studied. Half (50%) of the included studies had a low risk of bias. Three (30%) studies showed higher efficacy of botanical antifungals, five (50%) studies showed comparable results, and two (20%) studies showed higher efficacy of conventional antifungals in lesion improvement of oral candidiasis. The meta-analysis with random-effects analysis, which encompassed five studies involving 278 patients, revealed no significant difference in lesion improvement for oral candidiasis between botanical and conventional antifungals. The relative risk (RR) was calculated at 0.99, with a 95% confidence interval (CI) of (0.63, 1.56).ConclusionBased on the limited evidence, botanical antifungals have comparable efficacy to conventional antifungals in treating oral candidiasis. Therefore, they may serve as adjunctive or alternative treatments.Systematic Review Registrationhttps://www.crd.york.ac.uk/PROSPERO/view/CRD42024589391, identifier CRD42024589391.

Background and Purpose:Oral candidiasis is one of the most common fungal infections in humans. The treatment and prophylaxis of the patients suffering from this infection require the identification of new anti-Candida agents with no side effects or toxicity like medicinal plants. The present study was conducted to compare the antifungal activities of the aqueous, ethanolic, and methanolic extracts of the bark and roots of P. granatum with those of two routine antifungal agents (i.e., fluconazole and nystatin) on oral Candida strains isolated from liver transplant recipients.Materials and Methods:Minimum inhibitory concentrations (MICs) of the ethanolic, methanolic, and aqueous extracts of the bark and root of Punica granatum against C. albicans and C. glabrata isolated from oral cavities were evaluated according to the CLSI M27-A3. All data were analyzed in SPSS (version 16.0) by pairwise comparison and Kruskal-Wallis test.Results:The MIC50 and MIC90 values for the methanolic and ethanolic extracts of the bark and root of P. granatum against C. albicans were both obtained as 0.05 mg/ml with the geometric mean (GM) of 0.07. Furthermore, the MIC90 values for the aqueous extracts of bark and root were estimated as 0.05 and 0.2 mg/ml, respectively. With regard to C. glabrata, the MIC50 and MIC90 values for the methanolic and ethanolic extracts of the bark and root were 0.05 mg/ml. However, the MIC90 value for the aqueous extract against this species was obtained as 25 mg/ml. The GM values for the aqueous extracts of the bark and root were 9.49 and 0.32, respectively.Conclusion:As the findings indicated, the methanolic and ethanolic extracts of the bark and root of Punica granatum had anti-Candida activities. Therefore, they can be considered as mouthwash or toothpaste to prevent and treat Candida infections in the oral cavity.

ABSTRACTBackground and Aim:Canine pyoderma is a common dermatological condition, often caused by Staphylococcus pseudintermedius and related methicillin-resistant strains (MRSP and MRSS). Rising antimicrobial resistance necessitates alternative topical therapies. This study comparatively evaluated the in vitro antibacterial activity of P. betel leaf extract (both solvent-based and nanoemulsion forms) and cannabidiol (CBD) formulations against canine Staphylococcus isolates.Materials and Methods:Antibacterial activity was determined by broth microdilution to establish minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Five formulations were tested: Ethanolic betel leaf extract in dimethyl sulfoxide betel leaf (BL), BL extract nanoemulsion (BLN), CBD in ethanol, water-soluble CBD, and CBD nanoemulsion. Test organisms included 15 Staphylococcus isolates (five MRSP, five methicillin-susceptible S. pseudintermedius, and five MRSS) and five Pseudomonas aeruginosa. Gas chromatography-mass spectrometry (GC-MS) was used to characterize phytochemical constituents.Results:GC-MS revealed eugenol (40.86%) and hydroxychavicol (26.44%) as predominant antibacterial compounds. BL and BLN demonstrated potent anti-staphylococcal activity, with median MICs of 0.16 g/L and 0.31 g/L, respectively. BL exhibited significantly lower MIC and MBC values than BLN (p = 0.008). Among CBD formulations, ethanol-dissolved and water-soluble CBD displayed the strongest activity (median MICs 0.003 g/L and 0.004 g/L), while CBD nanoemulsion was markedly less effective (median MIC 7.50 g/L). BLN also exhibited antibacterial activity against P. aeruginosa (median MIC 0.62 g/L), comparable to BL.Conclusion:The novel BLN and soluble CBD formulations demonstrated significant in vitro antibacterial activity against methicillin-resistant Staphylococcus isolates from canine pyoderma. These results highlight their potential as topical antiseptic alternatives to chlorhexidine. Further in vivo studies are required to assess safety, efficacy, and formulation optimization. A combined betel-CBD nanoemulsion represents a promising direction for developing novel veterinary dermatological therapies.

Tooth decay is one of the most common chronic diseases in humans. The aim of this study was to investigate the inhibitory effects of Citrullus colocynthis extracts on the growth of Streptococcus mutans and Candida albicans and their cytotoxic effects on normal gingival fibroblast cells and breast cancer cells. The aqueous and alcoholic extracts of C. colocynthis fruit were evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, minimum inhibitory concentration (MICs) and minimal bactericidal concentration (MBCs)/minimal fungicidal concentration (MFCs) tests. The MICs and MBCs/MFCs were obtained from the aqueous extract (MIC 0.37 mg/mL and MBCs 1.5 mg/mL against S. mutans and MIC 0.37 mg/mL and MFCs 3.0 mg/mL against C. albicans) and ethanolic extract (MIC 0.75 mg/mL and MBCs 1.5 mg/mL against S. mutans and MIC 3.0 mg/mL and MFCs 12.0 mg/mL against C. albicans). The growth of S. mutans and C. albicans were effectively inhibited by extracts of C. colocynthis. The LC50 values of C. colocynthis on HGF1-PI cells were 4589.19 μg/mL and 3933.84 μg/mL by aqueous and ethanolic extracts, respectively. The LC50 values of C. colocynthis on MCF-7 cells were 4589.19 μg/mL and 3933.84 μg/mL by aqueous and ethanolic extracts, respectively. The extracts of C. colocynthis significantly decreased the growth of breast cancer and normal gingival fibroblast cells. The results of the study showed that the extracts may be used to treat oral mucosal diseases and prevent dental caries but future research is needed.

Candidal adherence has been implicated as the first step in the pathogenesis of oral candidosis, and germ tube formation by Candida albicans has been attributed as a co-factor that promotes adherence. Oral candidosis is treated with polyenes and the azole group of antifungal agents. As the intraoral concentrations of antifungals fluctuate considerably due to the dynamics of the oral cavity, we investigated the effect of short exposure to sub-lethal concentrations of antifungals on the germ tube formation of Candida albicans. After determining the minimum inhibitory concentration (MIC) of the antifungal agents, ten oral isolates of Candida albicans were exposed to sub-lethal concentrations of nystatin (6xMIC), amphotericin B (8xMIC), 5-fluorocytosine (8xMIC), ketoconazole (4xMIC) and fluconazole (4xMIC), for 1 h. Following removal of the antifungal agent and subsequent incubation in a germ tube-inducing medium, the germ tube formation of these isolates was quantified. When compared with the controls, exposure to nystatin and amphotericin B almost completely inhibited germ tube formation of all the isolates (mean percentage reduction of 97.68 and 97.52%, respectively; P<0.0001), while ketoconazole suppressed this activity to a lesser degree (30.84%; P=0.0174). However, 5-fluorocytosine- and fluconazole-mediated germ tube suppression was minimal (12.63 and 15.93%, respectively; P=0.3255 and P=0.3791). In clinical terms, these findings indicate that short exposure to sub-therapeutic levels of commonly prescribed antifungals may modulate candidal germ tube formation, and thereby the clearance of the organisms from the oral cavity.