Evaluation of glycyl-arginine and lysyl-aspartic acid dipeptides for their antimicrobial, antibiofilm, and anticancer potentials.

Antibacterial resistance and cancer are worldwide challenges and have been defined as major threats by international health organizations. Peptides are produced naturally by all organisms and have a variety of immunomodulatory, physiological, and wound-healing properties. They can also provide protection against microorganisms and tumor cells. Therefore, we aimed to determine the antimicrobial, antibiofilm, and anticancer potentials of Glycyl-Arginine and Lysyl-Aspartic acid dipeptides. The Broth Dilution and Crystal Violet Binding assays assessed the antimicrobial tests and biofilm inhibitory effects. The MTT assay was used to measure the cytotoxic effects of dipeptides on HeLa cell viability. According to our results, Candida tropicalis T26 and Proteus mirabilis U15 strains were determined as more resistant to Staphylococcus epidermidis W17 against Glycyl-Arginine and Lysyl-Aspartic acid dipeptides with MICs higher than 2 mM (1 mg/mL). Sub-MICs of Glycyl-Arginine caused inhibitions against biofilm formation of all the tested clinical isolates, with the highest inhibition observed against S. epidermidisW17. Lysyl-Aspartic acid exhibited zero to no effect against biofilm formation of P. mirabilisU15, and S. epidermidisW17, whereas it exhibited 52% inhibition of biofilm formation of C. tropicalisT26. Cell viability results revealed that HeLa cell viability decreases with increasing concentration of both dipeptides. Also, parallel to antimicrobial tests, Glycyl-Arginine has a greater cytotoxic effect compared to Lysyl-Aspartic acid. The findings from this study will contribute to the advancement of novel strategies involving dipeptide-based synthesizable molecules and drug development studies. However, it is essential to note that there are still challenges, including the need for extensive experimental and clinical trials.

Cervicovaginal Bacillus velezensis Isolate: A Potential Probiotic and an Antagonist Against Candida and Staphylococcus.

The cervicovaginal microbiota is an essential aspect of women's reproductive and overall health. In this study, we aimed to evaluate the probiotic properties of a cervicovaginal isolate, obtained from a gynecologically healthy woman and assess its antagonistic effect against various microorganisms isolated from the vagina. Cytological examination was performed using Papanicolaou staining, and the isolated microorganism was identified via 16S Ribosomal RNA Gene Sequence Analysis. Probiotic characteristics were evaluated by determining the tolerance of the isolate to low pH, different NaCl concentrations, and bile salts. Bacterial adherence to stainless steel sheets, antibiotic susceptibility, and antimicrobial activity tests were also conducted and analyzed. Antimicrobial tests and antagonistic activities were assessed through disc diffusion assays. The cervicovaginal isolate was identified as B. velezensis ON116948 and was found to be tolerant to low pH, high NaCl and 0.3% bile salts. Additionally, it exhibited adherence. With the exception of amoxicillin/clavulanic acid (AMC) (30 µg) and oxacillin (OX) (1 µg), this isolate was susceptible to all the antibiotics tested. Candida species did not grow on B. velezensis spread media, while B. velezensis was able to grow on C. albicans, C. glabrata, C. tropicalis, S. condimenti and S. epidermidis spread media with growth zones of 13.7 ± 0.6, 13.3 ± 0.6, 14.2 ± 4.4, 10.5 ± 0.5 and 16.0 ± 1.0 (around discs), respectively. Our findings suggest that the cervicovaginal B. velezensis ON116948 isolate exhibits probiotic properties and antagonistic activity. These results provide important insights into the potential use of this isolate as a probiotic for the prevention of vaginal infections.

Candida Biofilm Formation Assay on Essential Oil Coated Silicone Rubber.

Development of biofilm associated candidemia for patients with implanted biomaterials causes an urgency to develop antimicrobial and biofilm inhibitive coatings in the management of recalcitrant Candida infections. Recently, there is an increase in the number of patients with biofilm formation and resistance to antifungal therapy. Therefore, there is a growing interest to use essential oils as coating agents in order to prevent biomaterial-associated Candida infections. Often high costs, complicated and laborious technologies are used for both applying the coating and determination of the antibiofilm effects hampering a rapid screening of essential oils. In order to determine biofilm formation of Candida on essential oil coated surfaces easier, cheaper and faster, we developed an essential oil (lemongrass oil) coated surface (silicone-rubber) by using a hypromellose ointment/essential oil mixture. Furthermore, we modified the "crystal violet binding assay" to quantify the biofilm mass of Candida biofilm formed on the lemongrass oil coated silicone rubber surface. The essential oil coating and the biomass determination of biofilms on silicone rubber can be easily applied with simple and accessible equipment, and will therefore provide rapid information about whether or not a particular essential oil is antiseptic, also when it is used as a coating agent.

The relationship between the presence of HPV infection and biofilm formation in cervicovaginal smears.

PURPOSE: To demonstrate and understand the association of HPV infection and biofilm formation. METHODS: The study consisted of cervicovaginal samples of 72 women who were evaluated at the colposcopy unit. Papanicolaou staining was used for cytological examination while "Crystal Violet Binding" assay was performed to detect biofilm formation. RESULTS: HPV-DNA was positive in 55.5% (n = 40) of the patients. The biofilm formation rate was statistically significantly higher in the HPV-positive women (45%) compared to HPV-negative women (21.9%) (P < 0.05). There was a statistically significant relationship between the presence of single HPV and "high-risk HPV" types and biofilm formation (P < 0.05). Biofilm formation was found in 80% of women with abnormal smear demonstrating atypical epithelial cells (P < 0.05). CONCLUSION: Biofilm formation is more frequent at the cervicovaginal microbiota of patients with HPV infection. This finding is especially important in cases with atypical epithelial cells at their cervicovaginal smears.

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

ETHNOPHARMACOLOGICAL RELEVANCE: 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. AIM OF THE STUDY: 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. MATERIALS AND METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Electrospun essential oil-polycaprolactone nanofibers as antibiofilm surfaces against clinical Candida tropicalis isolates.

OBJECTIVE: As an approach to prevent biofilm infections caused by Candida tropicalis on various surfaces, determination of effect of biodegradable polycaprolactone nanofibers (PCLNFs) with different concentrations of two different essential oils were tested in this study. RESULTS: Both of the tested essential oils exhibited antifungal effect (minimal inhibitory concentration; 0.25-0.49 µL/mL, minimal fungicidal concentration; 0.25-0.49 µL/mL, depending on the C. tropicalis strain) (Zone of inhibition caused by 500 µL/mL concentration of oils; 28-56 mm). 0, 2, 4% clove oil PCLNFs and 0, 2, 4% red thyme oil-PCLNFs were free from bead formation and uniform in diameter. Diameters of all essential oil containing PCLNFs were ranged from 760 to 1100 nm and were significantly different from 0% essential oil-PCLNF (P < 0.05). 0, 2, 4% clove oil-PCLNFs were significantly more hydrophobic compared to 8% clove oil-PCLNF (P < 0.01), whereas 0% and 2% red thyme oil-PCLNFs were significantly more hydrophobic compared to 4% and 8% red thyme oil PCLNFs (P < 0.01). Highest amount of biofilm inhibition was observed by 4% clove oil-PCLNF and by 4% red thyme oil-PCLNF. CONCLUSIONS: Clove and red thyme oils may be used not only as antifungals but also as biofilm inhibitive agents on surfaces of biomaterials that are frequently contaminated by C. tropicalis, when they are incorporated into PCLNFs.

Distribution of clinical isolates of Candida spp. and antifungal susceptibility of high biofilm-forming Candida isolates

Abstract INTRODUCTION: The increase in the incidence of fungal infections, especially those caused by Candida albicans and other Candida species, necessitates the understanding and treatment of Candida-associated infections. In this study, we aimed to investigate the identification, distribution, and biofilm formation ability of different clinical Candida isolates and evaluate the distribution and antifungal susceptibilities of high biofilm-forming (HBF) Candida isolates. METHODS: For identification, carbohydrate fermentation, carbohydrate assimilation, and ChromAgar tests were used. Biofilm formation was assessed using crystal violet binding assay, while the susceptibility to antifungal agents was determined using ATBTM Fungus 3 test kits. RESULTS: The majority of Candida species were C. parapsilosis (31.3%; 31/99) and C. tropicalis (30.3%; 30/99). C. tropicalis was found to be the most frequently isolated species among all HBF Candida species. HBF Candida isolates were more frequently isolated from vaginal swab (35.7%; 10/28), tracheal aspirate (17.9%; 5/28), and urine (17.9%; 5/28). The majority of tested isolates were resistant to itraconazole and voriconazole, whereas no isolate was deemed resistant to 5-flucytosine. CONCLUSIONS: C. tropicalis displays the highest biofilm formation ability among all the Candida species evaluated, and HBF Candida isolates were more frequently seen in vaginal swab, tracheal aspirate, and urine samples. Our findings revealed that 5-flucytosine is the most efficient antifungal agent against HBF Candida isolates.

Multi drug resistance in strong biofilm forming clinical isolates of Staphylococcus epidermidis.

Staphylococcus epidermidis which exists in healthy human skin as a commensal inhabitant is also an important pathogen forming biofilms on many surfaces and recently, increased resistance traits were suggested to be acquired in biofilm environments. In this study; clinical Prevalences, antibiotic resistances and biofilm formations of S. epidermidis strains were determined and comparison of all these findings with each other was carried out in order to take precautions against them and figure out if high biofilm forming S. epidermidis strains display multi drug resistance. According to our results; samples of wound and blood were the most S. epidermidis isolated clinical materials (40%; 35%) and cardiothoracic surgery was the most S. epidermidis observed service unit. All of these strains were sensitive to vancomycin, however 65% of them showed resistance to all ß-lactam antibiotics (Penicillin, Oxacillin, Amoxicilin/Clavulonic acid), used in this study and 60% of all S. epidermidis strains were found as multi drug resistant. When the results of strong biofilm forming S. epidermidis strains are examined; they were isolated from sample of blood and service unit of cardiovascular surgery in highest frequency and 80% of them were ß-lactam resistant whereas 100% of them were multi drug resistant. One of these multi drug resistant strains which was resistant to maximum amount of different antimicrobial classes, was also observed as maximum biofilm forming strain among all the other S. epidermidis isolates. Multi drug resistance in strong biofilm forming strains shows that; biofilms play a role in antimicrobial resistance traits of S. epidermidis.

Multi drug resistance in strong biofilm forming clinical isolates of Staphylococcus epidermidis

Staphylococcus epidermidis which exists in healthy human skin as a commensal inhabitant is also an important pathogen forming biofilms on many surfaces and recently, increased resistance traits were suggested to be acquired in biofilm environments. In this study; clinical Prevalences, antibiotic resistances and biofilm formations of S. epidermidis strains were determined and comparison of all these findings with each other was carried out in order to take precautions against them and figure out if high biofilm forming S. epidermidis strains display multi drug resistance. According to our results; samples of wound and blood were the most S. epidermidis isolated clinical materials (40%; 35%) and cardiothoracic surgery was the most S. epidermidis observed service unit. All of these strains were sensitive to vancomycin, however 65% of them showed resistance to all β-lactam antibiotics (Penicillin, Oxacillin, Amoxicilin / Clavulonic acid), used in this study and 60% of all S. epidermidis strains were found as multi drug resistant. When the results of strong biofilm forming S. epidermidis strains are examined; they were isolated from sample of blood and service unit of cardiovascular surgery in highest frequency and 80% of them were β-lactam resistant whereas 100% of them were multi drug resistant. One of these multi drug resistant strains which was resistant to maximum amount of different antimicrobial classes, was also observed as maximum biofilm forming strain among all the other S. epidermidis isolates. Multi drug resistance in strong biofilm forming strains shows that; biofilms play a role in antimicrobial resistance traits of S. epidermidis.