Clonal Diversity, Antimicrobial Susceptibility and Presence of Genes Encoding Virulence Factors in Staphylococcus aureus Strains Isolated from Cut Wound Infections.

The aim of the study was to evaluate the clonal relatedness and antimicrobial susceptibility in 52 Staphylococcus aureus strains isolated from cut wound infections in non-related community patients and to determine the presence of selected virulence genes. To analyse the clonal relatedness of investigated strains, pulsed-field gel electrophoresis (PFGE) of macrorestricted DNA fragments was conducted. Antimicrobial susceptibility testing was performed using the AST-P644 card in the VITEK 2 Compact system. All strains were tested for the presence of selected virulence genes using Single and Multiplex PCR. All isolates were classified into 15 PFGE genotypes and seven unique patterns. The vast majority of investigated S. aureus strains were susceptible to all tested antimicrobial agents. Among examined S. aureus strains, 24 combinations of virulence factors were identified. 62.5% of S. aureus strains contained various egc types, alone or together with other staphylococcal enterotoxin genes. A high percentage (86.5%) of isolates harboured superantigen genes. The most frequent enterotoxin gene identified was encoding for sep. All S. aureus strains were classified as agr-positive, and the most frequent agr gene was agr-1. Our results indicate that all examined strains isolated from cut wound infections demonstrated high clonal diversity, diversified gene distribution and good susceptibility to antimicrobial agents.

Changes in Growth, Ionic Status, Metabolites Content and Antioxidant Activity of Two Ferns Exposed to Shade, Full Sunlight, and Salinity.

The interactions between ferns and the environment have been frequently researched. However, detailed data on how ferns respond to specific stresses and a combination of stress factors during cultivation are lacking. This study assessed the effects of salinity and full sunlight and the combination of both stresses on the growth and selected metabolic parameters of two hardy ferns (Athyrium nipponicum cv. Red Beauty and Dryopteris erythrosora) under production conditions. Hardy ferns are highly interesting ornamental plants that can serve as a potential source of antioxidants for the pharmaceutical, cosmetic, and food industries. The results showed that in both ferns, salinity and salinity combined with full sunlight lowered the dry weight of the aerial part and potassium/sodium and calcium/potassium ratio compared with control plants. Salinity, full sunlight, and multi-stress did not affect the total polyphenol content in both ferns but increased the total free amino acids and flavonoids in D. erythrosora. In A. nipponicum cv. Red Beauty, all stressors decreased the total free amino acids content and the antioxidant activities determined by ABTS, DPPH, FRAP, and reducing power assays. By contrast, plants of D. erythrosora grown under full sunlight are characterized by higher antioxidant activities determined by DPPH, FRAP, and reducing power assays. Overall, a greater adaptive potential to abiotic stresses was found in D. erythrosora than in A. nipponicum cv. Red Beauty. Our findings shed some light on the physiological mechanisms responsible for sensitivity/tolerance to salinity, full sunlight, and combined stresses in hardy ferns.

Bioactive Carboxymethyl Cellulose (CMC)-Based Films Modified with Melanin and Silver Nanoparticles (AgNPs)-The Effect of the Degree of CMC Substitution on the In Situ Synthesis of AgNPs and Films' Functional Properties.

Green synthesis of nanoparticles for use in food packaging or biomedical applications is attracting increasing interest. In this study, the effect of the degree of substitution (0.7, 0.9 and 1.2) of a carboxymethylcellulose polymer matrix on the synthesis and properties of silver nanoparticles using melanin as a reductant was investigated. For this purpose, the mechanical, UV-Vis barrier, crystallinity, morphology, antioxidant and antimicrobial properties of the films were determined, as well as the color and changes in chemical bonds. The degree of substitution effected noticeable changes in the color of the films (the L* parameter was 2.87 ± 0.76, 5.59 ± 1.30 and 13.45 ± 1.11 for CMC 0.7 + Ag, CMC 0.9 + Ag and CMC 1.2 + Ag samples, respectively), the UV-Vis barrier properties (the transmittance at 280 nm was 4.51 ± 0.58, 7.65 ± 0.84 and 7.98 ± 0.75 for CMC 0.7 + Ag, CMC 0.9 + Ag and CMC 1.2 + Ag, respectively) or the antimicrobial properties of the films (the higher the degree of substitution, the better the antimicrobial properties of the silver nanoparticle-modified films). The differences in the properties of films with silver nanoparticles synthesized in situ might be linked to the increasing dispersion of silver nanoparticles as the degree of CMC substitution increases. Potentially, such films could be used in food packaging or biomedical applications.

Regulatory and Enterotoxin Gene Expression and Enterotoxins Production in Staphylococcus aureus FRI913 Cultures Exposed to a Rotating Magnetic Field and trans-Anethole.

The study aimed to examine the influence of a rotating magnetic field (RMF) of two different frequencies (5 and 50 Hz) on the expression of regulatory (agrA, hld, rot) and staphylococcal enterotoxin (SE-sea, sec, sel) genes as well as the production of SEs (SEA, SEC, SEL) by the Staphylococcus aureus FRI913 strain cultured on a medium supplemented with a subinhibitory concentration of trans-anethole (TA). Furthermore, a theoretical model of interactions between the bacterial medium and bacterial cells exposed to RMF was proposed. Gene expression and SEs production were measured using quantitative real-time PCR and ELISA techniques, respectively. Based on the obtained results, it was found that there were no significant differences in the expression of regulatory and SE genes in bacteria simultaneously cultured on a medium supplemented with TA and exposed to RMF at the same time in comparison to the control (unexposed to TA and RMF). In contrast, when the bacteria were cultured on a medium supplemented with TA but were not exposed to RMF or when they were exposed to RMF of 50 Hz (but not to TA), a significant increase in agrA and sea transcripts as compared to the unexposed control was found. Moreover, the decreased level of sec transcripts in bacteria cultured without TA but exposed to RMF of 50 Hz was also revealed. In turn, a significant increase in SEA and decrease in SEC and SEL production was observed in bacteria cultured on a medium supplemented with TA and simultaneously exposed to RMFs. It can be concluded, that depending on SE and regulatory genes expression as well as production of SEs, the effect exerted by the RMF and TA may be positive (i.e., manifests as the increase in SEs and/or regulatory gene expression of SEs production) or negative (i.e., manifests as the reduction in both aforementioned features) or none.

Novel Gluten-Free Bread with an Extract from Flaxseed By-Product: The Relationship between Water Replacement Level and Nutritional Value, Antioxidant Properties, and Sensory Quality.

The food industry generates a great amount of food waste and by-products, which in many cases are not fully valorized. Press cakes, deriving from oilseeds extraction, represent interesting co-products due to their nutritional value, high biopolymers content, and the presence of bioactive phytochemicals. Gluten-free breads (GFBs) are products that have disadvantages such as unsatisfactory texture, low nutritional value, and short shelf life, so natural additives containing proteins and hydrocolloids are in demand to increase GFBs value. In this study, extract from flaxseed by-product (FOCE-Flaxseed Oil Cake Extract) was used to replace water (25-100%) in GFBs formulations and their nutritional value, antioxidant properties, and sensory features were investigated. The results showed that GFBs with FOCE had an elevated nutritional and nutraceutical profile (up to 60% more proteins, significantly increased K, Mg, and P levels). Moreover, the addition of FOCE improved the technological parameters (increased specific volume, number of cells and height/width ratio, reduced density, average size, and perimeter of cells), antioxidant potential, and overall sensory quality of GFBs. This study showed an encouraging way of using a by-product that, due to its high content of proteins, polysaccharides, minerals, and antioxidants, can add value to GFBs.

Inhibitory Effect of Eugenol and trans-Anethole Alone and in Combination with Antifungal Medicines on Candida albicans Clinical Isolates.

One of the most common pathogens among yeasts is Candida albicans, which presents a serious health threat. The study aimed to check the antifungal properties of trans-anethole and eugenol with selected antifungal medicines (AMs) against C. albicans clinical isolates. The checkerboard method was used to tests of interactions between these compounds. Achieved results indicated that eugenol showed synergistic and additive activities with miconazole and econazole against investigated clinical isolates, respectively. Moreover, the combination - trans-anethole - miconazole also showed an additive effect against two clinical isolate. We tried to relate the results to changes in C. albicans cell sheaths under the influence of essential oils compounds (EOCs) performing the Fourier transform infrared spectra analysis to confirm the presence of particular chemical moieties in C. albicans cells. Nevertheless, no strong relationships was observed between synergistic and additive actions of used EOC-AMs combinations and chemical moieties in C. albicans cells.

Evaluation of Chemical Changes in Laboratory-Induced Colistin-Resistant Klebsiella pneumoniae.

This study evaluates the electrical potential and chemical alterations in laboratory-induced colistin-resistant Klebsiella pneumoniae, as compared to the susceptible strain using spectroscopic analyses. The minimal inhibitory concentration (MIC) of colistin, ζ-potential and chemical composition analysis of K. pneumoniae strains are determined. The results obtained for the K. pneumoniaeCol-R with induced high-level colistin resistance (MIC = 16.0 ± 0.0 mg/L) are compared with the K. pneumoniaeCol-S strain susceptible to colistin (MIC = 0.25 ± 0.0 mg/L). Fourier transform infrared (FTIR) and Raman spectroscopic studies revealed differences in bacterial cell wall structures and lipopolysaccharide (LPS) of K. pneumoniaeCol-R and K. pneumoniaeCol-S strains. In the beginning, we assumed that the obtained results could relate to a negative charge of the bacterial surface and different electrostatic interactions with cationic antibiotic molecules, reducing the affinity of colistin and leading to its lower penetration into K. pneumoniaeCol-R cell. However, no significant differences in the ζ-potential between the K. pneumoniaeCol-R and K. pneumoniaeCol-S strains are noticed. In conclusion, this mechanism is most probably associated with recognisable changes in the chemical composition of the K. pneumoniaeCol-R cell wall (especially in LPS) when compared to the susceptible strain.

Antioxidant Potential of Flower Extracts from Centaurea spp. Depends on Their Content of Phenolics, Flavonoids and Free Amino Acids.

Scientists intensely search for new sources of antioxidants, perceived as important health-promoting agents. Some species of the large genus Centaurea provide raw materials for the pharmaceutical and cosmetic industries, as well as produce edible flowers. This is the first study that determines the content of total polyphenols, flavonoids, reducing sugars, free amino acids and the antioxidant potential in the flower extracts of C. nigra L., C. orientalis L. and C. phrygia L. The total polyphenol and flavonoid content is the highest in the extract of C. orientalis, and the lowest in that of C. phrygia. Similarly, C. orientalis shows the greatest scavenging activity on DPPH (1,1-diphenyl-2-picryl-hydrazyl), ABTS [2,2'-azobis(3-ethylbenzothiazoline-6-sulfonate)] and Fe3+ reducing power assays, whereas the lowest activity is found for C. phrygia. The highest content of reducing sugars is found in C. nigra, while C. orientalis has the highest levels of free amino acids. We find a strong positive correlation between total phenolics and flavonoids and the antioxidant capacity of all three Centaurea species. Moreover, the content of free amino acids strongly and positively correlates with the levels of total phenolics and flavonoids, antioxidant activity assessed by DPPH and ABTS assays and Fe3+ reducing power. Summing up, C. orientalis exhibits the strongest antioxidant potential of the investigated Centaurea species. This species could potentially be a natural source of antioxidant substances for the pharmacy, cosmetics and food industries. The content of free amino acids may be used as a marker of the antioxidant status of Centaurea species.

Antibacterial Activity of Selected Essential Oil Compounds Alone and in Combination with ß-Lactam Antibiotics Against MRSA Strains.

This study aimed to determine the effect of selected essential oil compounds (EOCs) on the antibacterial activity of ß-lactam antibiotics (ßLAs) against methicillin-resistant Staphylococcus aureus (MRSA) strains. The following parameters were studied: antibiotic susceptibility testing, detection of mecA gene and evaluation of genotypic relativity of isolates using molecular techniques, analysis of chemical composition applying Fourier-transform infrared (FTIR) spectroscopy, and determination of antibacterial activity of EOCs alone and in combination with ßLAs against MRSA strains using microdilution and checkerboard methods. It was found that all isolates expressed MRSA and resistance phenotypes for macrolides, lincosamides, and streptogramins B. All isolates harbored the mecA gene and belonged to three distinct genotypes. Eight of the 10 EOCs showed efficient antimicrobial activity against the MRSA reference strain. The analysis of interaction between EOCs and ßLAs against the MRSA reference strain revealed a synergistic and additive effect of the following combinations: methicillin (Met)-linalyl acetate (LinAc), penicillin G (Pen)-1,8-cineole (Cin), and Pen-LinAc. Analysis of EOC-ßLA interactions showed a synergistic and additive effect in the following combinations: Met-LinAc (against low- and high-level ßLAs resistance strains), Pen-Cin, and Pen-LinAc (against low-level ßLAs resistance strains). It was also confirmed that changes in phosphodiester, -OH, -CH2 and -CH3 groups may change the interactions with ßLAs. Moreover, the presence of two CH3O- moieties in the Met molecule could also play a key role in the synergistic and additive mechanism of LinAc action with Met against MRSA strains. Direct therapy using a Met-LinAc combination may become an alternative treatment method for staphylococcal infections caused by MRSA. However, this unconventional therapy must be preceded by numerous cytotoxicity tests.

Innate Immune Response against Staphylococcus aureus Preincubated with Subinhibitory Concentration of trans-Anethole.

The study aimed to analyze morphological and functional changes of Staphylococcus aureus cells due to trans-anethole (a terpenoid and the major constituent of fennel, anise, or star anise essential oils) exposition, and their consequences for human neutrophils phagocytic activity as well as IL-8 production (recognized as the major chemoattractant). The investigation included the evaluation of changes occurring in S. aureus cultures, i.e., staphyloxanthin production, antioxidant activities, cell size distribution, and cells composition as a result of incubation with trans-anethole. It was found that the presence of trans-anethole in the culture medium reduced the level of staphyloxanthin production, as well as decreased antioxidant activities. Furthermore, trans-anethole-treated cells were characterized by larger size and a tendency to diffuse in comparison to the non-treated cells. Several cell components, such as phospholipids and peptidoglycan, were found remarkably elevated in the cultures treated with trans-anethole. As a result of the aforementioned cellular changes, the bacteria were phagocytized by neutrophils more efficiently (ingestion and parameters associated with killing activity were at a higher level as compared to the control system). Additionally, IL-8 production was at a higher level for trans-anethole modified bacteria. Our results suggest that trans-anethole represents a promising measure in combating severe staphylococcal infections, which has an important translational potential for clinical applications.

The Effect of Fermentation with Kefir Grains on the Physicochemical and Antioxidant Properties of Beverages from Blue Lupin (Lupinus angustifolius L.) Seeds.

Plant derived fermented beverages have recently gained consumers' interest, particularly due to their intrinsic functional properties and presence of beneficial microorganisms. Three variants containing 5%, 10%, and 15% (w/w) of sweet blue lupin (Lupinus angustifolius L. cv. "Boregine") seeds were inoculated with kefir grains and incubated at 25 °C for 24 h. After processing, beverages were stored in refrigerated conditions (6 °C) for 21 days. Changes in microbial population, pH, bioactive compounds (polyphenolics, flavonoids, ascorbic acid), reducing sugars, and free amino acids were estimated. Additionally, viscosity, firmness, color, and free radicals scavenging properties were determined. Results showed that lactic acid bacteria as well as yeast were capable of growing well in the lupin matrix without any supplementation. During the process of refrigeration, the viability of the microorganisms was over the recommended minimum level for kefir products. Hydrolysis of polysaccharides as well as increase of free amino acids was observed. As a result of fermentation, the beverages showed excellent DPPH, ABTS+·, ·OH, and O2- radicals scavenging activities with a potential when considering diseases associated with oxidative stress. This beverages could be used as a new, non-dairy vehicle for beneficial microflora consumption, especially by vegans and lactose-intolerant consumers.

The Influence of Essential Oil Compounds on Antibacterial Activity of Mupirocin-Susceptible and Induced Low-Level Mupirocin-Resistant MRSA Strains.

Because of the bacterial drug resistance development, it is reasonable to investigate chemical compounds capable of preventing the spread of resistance to mupirocin (MUP), commonly used in staphylococcal eradication. The objective of the study was to verify the influence of essential oil compounds (EOCs) on the antibacterial activity of MUP against mupirocin-susceptible (MupS) and induced low-level mupirocin-resistant (MupRL) methicillin-resistant Staphylococcus aureus (MRSA) strains. The following parameters were examined: MRSAMupS and MRSAMupRL susceptibility to EOCs (1,8-cineole, eugenol, carvacrol, linalool, (-)-menthone, linalyl acetate, and trans-anethole), the bacterial cell size distribution, and chemical composition by the use of Fourier Transform Infrared Spectroscopy (FTIR) and Raman spectroscopies. The MRSAMupS and MRSAMupRL strains were susceptible to all tested EOCs. 1,8-cineole and (-)-menthone showed synergistic activity against MRSAMupS in combination with mupirocin, whereas 1,8-cineole exhibited synergistic activity against MRSAMupRL as well. In-depth analysis showed that both MRSAMupS and MRSAMupRL displayed similar distributions of the bacterial cell size. The FTIR and Raman spectra of the MRSAMupS and MRSAMupRL strains showed differences in some regions. New bands in the MRSAMupRL Raman spectrum were observed. It was concluded that the use of 1,8-cineole in combination with mupirocin can increase the mupirocin activity against the MRSAMupS and MRSAMupRL strains.

The Antibacterial Activity of Lavender Essential Oil Alone and In Combination with Octenidine Dihydrochloride against MRSA Strains.

In the post-antibiotic era the issue of bacterial resistance refers not only to antibiotics themselves but also to common antiseptics like octenidine dihydrochloride (OCT). This appears as an emerging challenge in terms of preventing staphylococcal infections, which are both potentially severe and easy to transfer horizontally. Essential oils have shown synergisms both with antibiotics and antiseptics. Therefore the aim of this study was to investigate the impact of lavender essential oil (LEO) on OCT efficiency towards methicillin-resistant S. aureus strains (MRSA). The LEO analyzed in this study increased the OCT's susceptibility against MRSA strains. Subsequent FTIR analysis revealed cellular wall modifications in MRSA strain cultured in media supplemented with OCT or LEO/OCT. In conclusion, LEO appears to be a promising candidate for an efficient enhancer of conventional antiseptics.

The Influence of Accelerated UV-A and Q-SUN Irradiation on the Antimicrobial Properties of Coatings Containing ZnO Nanoparticles.

The influence of accelerated UV-A and Q-SUN irradiation on the antimicrobial properties of coatings containing ZnO nanoparticles was investigated using a polyethylene (PE) film covering. The results of the study showed that Methyl Hydroxypropyl Celluloses (MHPC) coatings did not influence the growth of S. aureus, B. cereus, E. coli, P. aeruginosa or C. albicans cells. MHPC coatings containing ZnO nanoparticles inhibited the growth of bacterial strains and reduced the number of C. albicans strains. Accelerated Q-SUN and UV-A irradiation had no influence on the antimicrobial effect of nano ZnO coatings against S. aureus, B. cereus and E. coli. Q-SUN irradiation decreased the activity of MHPC coatings containing nanoparticles against P. aeruginosa and C. albicans. An FT-IR analysis clearly showed that ZnO nanoparticles shielded the MHPC coating during Q-SUN irradiation.

Spray-Drying Impact the Physicochemical Properties and Formation of Maillard Reaction Products Contributing to Antioxidant Activity of Camelina Press Cake Extract.

Spray-drying is one of the most popular techniques in the food industry for converting liquid material from a fluid state into a form of dried particles to produce encapsulated or instant products. Instant products are considered as convenient foods; moreover, the goal of encapsulation is to close the bioactive compounds in a shell, preventing them from being affected by environmental factors. The purpose of this study was to examine the influence of spray-drying conditions, in particular three inlet temperatures, on the physicochemical and antioxidant properties of powders obtained from Camelina Press Cake Extract (CPE). The CPE was spray-dried at 140 °C, 160 °C and 180 °C. The solubility, Carr and Hausner Indexes, tapped densities and water activity of the powders were analyzed. The structural changes were also detected using FTIR spectroscopy. Additionally, the characteristics of the initial and reconstituted samples and their rheological properties were evaluated. The antioxidant potential, total polyphenols and flavonoids content, free amino acids, and the Maillard reaction products contents in the spray-dried powders were also evaluated. The results indicate a cascade of changes between the initial and reconstituted samples, and important changes in the bioactive potential of samples. The inlet temperature significantly influenced the solubility, flowability and particle sizes of the powders, as well as Maillard products formation. The results of the rheological measurements illustrate the changes after the reconstitution of extracts. This study indicates the optimal parameters of CPE spray-drying, those that yield favorable physicochemical and functional values, which may open up a promising path for CPE valorization, indicating its potential and the possibilities of its use.

Fermented Tea as a Food with Functional Value-Its Microbiological Profile, Antioxidant Potential and Phytochemical Composition.

Kombucha is a fermented tea drink produced by a symbiotic culture of bacteria and yeast, known as SCOBY. Its base has traditionally been black tea, which has been recognized for its health-promoting properties, particularly its antioxidant activity based on its high content of pol-yphenolic compounds. A number of previous studies have demonstrated the equally favourable biochemical and phytochemical composition of green tea. The aim of this study was to analyse and compare the basic biochemical composition, microbiological composition and antioxidant properties of black and green tea-based Kombucha. The green tea-based Kombucha showed a quantitatively more abundant microbial composition (Lactic Acid Bacteria, Acetobacter sp., Yeast), a higher reducing potential (FRAP-4326.58 Fe(II)µM/L) and a higher content of total polyphenols (23.84 mg GAE/100 mL, reducing sugars (3212.00 mg/100 mL) as well as free amino acids (849.00 mg GLY/mL). Kombucha made from black tea, on the other hand, showed a higher anti-oxidant potential (1.17 Trolox (mM) TEAC), neutralising the DPPH radical at 94.33% and ABTS at 97.74%. It also had a higher level of acetic acid (0.08 g/100 mL). Green tea kombucha had a higher scavenging capacity of 90.6% for superoxide radical (O2-) and 69.28% for hydroxyl radical (·OH) than black tea kombucha. In the present study, both kombucha drinks tested were shown to be source of potent antioxidants. In addition, green tea, as a kombucha base, has proven to be as beneficial a raw material that will provide full nutritional and health-promoting values as traditional kombucha.

Effect of Flaxseed Oil Cake Extract on the Microbial Quality, Texture and Shelf Life of Gluten-Free Bread.

Extending the shelf life of gluten-free bread (GFB) is a challenge. Mainly due to the ingredients used and their characteristics, GFB has numerous drawbacks such as unsatisfactory texture and rapid staling beyond a low nutritional value. In the present study, flaxseed oil cake extract (FOCE) was used to replace water (25-100%) in GFB formulations in order to test FOCE's potential to reduce GFB staling and extend microbial stability. Texture (TPA test), water activity (LF NMR), acidity (pH measurements) and microbiological quality of GFBs were tested. Moreover, the content of a lignan with broad health-promoting potential, secoisolariciresinol diglucoside (SDG), in GFB with FOCE was analyzed. The results showed that the use of FOCE enriched experimental GFB in valuable SDG (217-525 µg/100 g DM) while not causing adverse microbiological changes. A moderate level (25-50%) of FOCE did not change the main texture parameters of GFB stored for 72 h, the quality of which was comparable to control bread without FOCE. Meanwhile, higher proportions of FOCE (75-100% of water replacement) shortened GFB shelf life as determined by water activity and texture profile, suggesting that GFB with FOCE should be consumed fresh. To summarize, FOCE at moderate levels can add value to GFBs without causing a drop in quality, while still fitting in with the idea of zero waste and the circular economy.

Recent progress in pectin extraction, characterization, and pectin-based films for active food packaging applications: A review.

Pectin is an abundant complex polysaccharide obtained from various plants. Safe, biodegradable, and edible pectin has been extensively utilized in the food industry as a gelling agent, thickener, and colloid stabilizer. Pectin can be extracted in a variety of ways, thus affecting its structure and properties. Pectin's excellent physicochemical properties make it suitable for many applications, including food packaging. Recently, pectin has been spotlighted as a promising biomaterial for manufacturing bio-based sustainable packaging films and coatings. Functional pectin-based composite films and coatings are useful for active food packaging applications. This review discusses pectin and its use in active food packaging applications. First, basic information and characteristics of pectin, such as the source, extraction method, and structural characteristics, were described. Then, various methods of pectin modification were discussed, and the following section briefly described pectin's physicochemical properties and applications in the food sector. Finally, the recent development of pectin-based food packaging films and coatings and their use in food packaging were comprehensively discussed.

Improved efficacy of eugenol and trans-anethole in combination with octenidine dihydrochloride against Candida albicans and Candida parapsilosis.

INTRODUCTION AND OBJECTIVE: Candidiasis is a fungal infection caused by yeasts from the Ogenus Candida. Considering increasing antifungal resistance rates the activity was analyzed of natural compounds to eradicate Candida spp. The aim of the study was to check the antifungal activity of selected essential oil compounds (EOCs; thymol, menthol, eugenol [E], carvacrol, trans-anethole [TA]) alone, and in combination with octenidine dihydrochloride (OCT) against C. albicans and C. parapsilosis reference, and clinical strains. MATERIAL AND METHODS: Investigated clinical isolates were obtained from skin wounds of patients treated for superficial wounds candidiasis. The following parameters were studied: antifungal susceptibility testing using the VITEK system, antifungal activity of EOCs alone and in combination with OCT using microdilution and checkerboard assays, antifungal efficacy of selected chemicals using time-kill curve assay, and changes in cell permeability in the presence of selected chemicals using crystal violet assay. RESULTS: Clinical isolates of C. albicans and C. parapsilosis were resistant to fluconazole and voriconazole. The highest inhibition activity against Candida isolates was observed for E. The OCT - TA and OCT - E combinations showed synergistic and additive activities against all strains, respectively. These combinations also appeared to affect the rate of yeast cell killing and increasing the permeability of Candida cells. CONCLUSIONS: The study indicates that E and TA potentially used in formulation with OCT might eradicate pathogenic yeasts; however, microbiological and clinical studies are still required.

Effect of Selected Entomopathogenic Fungal Species on Embryonic Development of Ascaris suum (Nematoda).

The aim of the study was to evaluate the potential of using five selected species of entomopathogenic fungi (Beauveria bassiana, B. brongniartii, Conidiobolus coronatus, Isaria fumosorosea, and Metarhizium robertsii) in the bioregulation of the dispersive stages of the parasitic nematode-Ascaris suum. Experimental cultures of each of the selected entomopathogenic fungi, as well as a control culture without fungi, were incubated with A. suum eggs at 26 °C for 28 days. Development of the A. suum eggs was observed using a light microscope on the 7th, 14th, 21st, and 28th days of incubation. The API-ZYM® test was used to determine, semiquantitatively, the activity of 19 hydrolytic enzymes from the entomopathogenic fungi. The cytotoxicity of the fungi was determined using tetrazole salt MTT. It was found that none of the five tested strains of entomopathogenic fungi showed an ovicidal effect, and none of them colonized the A. suum egg shells. However, ovistatic activity was observed mainly until the 14th day of incubation by I. fumosorosea, M. robertsii, and B. bassiana. In the MTT test, M. robertsii showed moderate cytotoxicity, while the other species showed low cytotoxicity. Among the strains tested, I. fumosorosea showed the highest spectrum of hydrolase production (13 out of 19 enzymes gave a positive reaction from 3 to 5; 20-40 nM or more). The absence of morphological changes in the A. suum egg shells suggests that the antagonistic effect of the studied entomopathogenic fungi may be due to their cytotoxicity, associated with the production of secondary metabolites-toxins (M. robertsii) and enzymatic activity (I. fumosorosea).