Resveratrol/Selenium Nanocomposite with Antioxidative and Antibacterial Properties.

In this work, we synthesized a new composite material comprised of previously formulated resveratrol nanobelt-like particles (ResNPs) and selenium nanoparticles (SeNPs), namely ResSeNPs. Characterization was provided by FESEM and optical microscopy, as well as by UV-Vis and FTIR spectroscopy, the last showing hydrogen bonds between ResNPs and SeNPs. DPPH, TBA, and FRAP assays showed excellent antioxidative abilities with ResNPs and SeNPs contributing mainly to lipid peroxidation inhibition and reducing/scavenging activity, respectively. The antibacterial effect against common medicinal implant colonizers pointed to notably higher activity against Staphylococcus isolates (minimal inhibitory concentrations 0.75-1.5%) compared to tested gram-negative species (Escherichia coli and Pseudomonas aeruginosa). Antibiofilm activity against S. aureus, S. epidermidis, and P. aeruginosa determined in a crystal violet assay was promising (up to 69%), but monitoring of selected biofilm-related gene expression (pelA and algD) indicated the necessity of the involvement of a larger number of genes in the analysis in order to further establish the underlying mechanism. Although biocompatibility screening showed some cytotoxicity and genotoxicity in MTT and alkaline comet assays, respectively, it is important to note that active antioxidative and antibacterial/antibiofilm concentrations were non-cytotoxic and non-genotoxic in normal MRC-5 cells. These results encourage further composite improvements and investigation in order to adapt it for specific biomedical purposes.

Could alder buckthorn (Frangula alnus Mill) be a source of chemotherapeutics effective against hepato- and colorectal carcinoma? An in vitro study.

Among numerous types of cancer, hepatocellular and colorectal carcinoma are important causes of mortality. Given the nature of these cancer types and their resistance, it is of great importance to find new chemotherapeutics and therapy targets, so plant products seem to be an excellent choice in such search. The main goal of this study was to investigate anticancer activity of Frangula alnus ethyl-acetate extract (FA) and its dominant constituent emodin (E) on hepatocellular and colorectal carcinoma cell lines, HepG2 and HCT116, as well as on normal MRC-5 fibroblasts. Cytotoxicity was investigated in MTT test and both FA and E showed strong reduction of cell viability in cancer cells. Flow cytometer analysis demonstrated that FA and E led to G1 phase arrest and slight accumulation of cells in the G2/M phase; additionally, annexinV-FITC/7AAD dying showed that FA and E decreased cell viability and triggered apoptosis in all cell lines. FA and E evidenced strong genotoxic potential in comet assay performed on all cell lines, while tests measuring antioxidative potential (DPPH and TBA) demonstrated strong effect of FA. It could be concluded that both FA and E have significant anticancer activity against hepatocellular and colorectal carcinoma cell lines HepG2 and HCT116, but notable selectivity was not observed.

Cuticular Wax Variability of Abies alba, A.×borisii-regis and A.†cephalonica from the Balkans: Chemophenetic and Ecological Aspects.

This is the first study on cuticular wax variability of Abies alba, A.×borisii-regis and A. cephalonica, using 18 native populations from the assumed hybrid zone in the Balkan Peninsula. Presence of 13 n-alkanes with chain-lengths ranging from C21 to C33 , one primary alcohol, two diterpenes, one triterpene and one sterol was determined in hexane extracts of 269 needle samples. The multivariate statistical analyses at the population level entirely failed in supporting circumscription of Balkan Abies taxa and therefore, in identifying hybrid populations. However, performed at the species level, these analyses revealed a certain tendency of differentiation between A. alba and A. cephalonica, while individuals of A.×borisii-regis were largely overlapped by the clouds of both parent species. Finally, the correlation analysis suggested that the observed variation of wax compounds was probably genetically conditioned and that it does not represent an adaptive response to various environmental factors.

The role of Gentiana lutea extracts in reducing UV-induced DNA damage.

Ultraviolet (UV) radiation can result in DNA damage, mainly through direct formation of pyrimidine dimers and generation of reactive oxygen species, which can lead to the skin disorders including cancer. In accordance with this, the use of natural antigenotoxins and/or antioxidants could contribute to human health protection. Considering that plants are rich in both, the aim of this study was to investigate UV-protective and antioxidative properties of yellow gentian (Gentiana lutea), being well established in pharmacopeias and traditional medicine. Tested extracts were derived from root and shoot of the in vitro cultivated plants. Prescreening of the genotoxic properties of UVC, UVA, and the extracts, as well as the extracts' antigenotoxicity were estimated by applying alkaline comet assay on normal fetal lung fibroblast (MRC-5) and human melanoma cells (Hs 294T). Antioxidant potential was tested in ferrous ions chelating ferric reducing antioxidant power and cupric reducing antioxidant capacity assays. Genotoxicity testing, which revealed moderate DNA-damaging potential of root extract on MRC-5 cells and high genotoxicity of shoot extract on both cell lines, pointed out nongenotoxic concentrations that could be used in antigenotoxicity assay. Doses of 63 and 3 J/cm2 for UVC and UVA, respectively, were established for antigenotoxicity study, since they induced sufficient DNA damage without notable cytotoxicity. Results of antigenotoxicity revealed strong protective effect of both extracts against UVC (the highest inhibitions 58% and 47%) and UVA (the highest inhibitions 69% and 60%), in Hs 294T and MRC-5 cells, respectively. Study of the antioxidative properties demonstrated stronger activity of shoot extract. Results obtained proved to be encouraging but further research of the UV-protective role of Gentiana lutea extracts and underlying molecular mechanisms is recommended.

Essential oils as adjuvants in endodontic therapy: myth or reality?

To enhance endodontics, the search for new antibacterials that can improve infected tooth root canal treatment is ongoing. As potent antibacterial and antibiofilm agents, essential oils (EOs) have been suggested as novel endodontic materials. Several studies indicate that EO-based irrigants and medicaments show promising reductive potential against the most important intracanal pathogen, Enterococcus faecalis, and notably contribute to intracanal biofilm eradication. In terms of additional benefits that EO-based endodontic materials can provide, their antioxidant and anti-inflammatory potential are also important, but they have only scarcely been explored in research. Investigations into the benefits of EO-based endodontic materials together with their biocompatibility are needed. The results presented in this review strongly encourage further research on this topic.

Hesperetin-Between the Ability to Diminish Mono- and Polymicrobial Biofilms and Toxicity.

Hesperetin is the aglycone of citrus flavonoid hesperidin. Due to the limited information regarding hesperetin antimicrobial potential and emerging need for novel antimicrobials, we have studied its antimicrobial activity (microdilution assay), antibiofilm activity with different assays in two models (mono- and polymicrobial biofilm), and toxicity (MTT and brine shrimp lethality assays). Hesperetin inhibited growth of all Candida isolates (minimal inhibitory concentration, MIC, 0.165 mg/mL), while it's inhibitory potential towards Staphylococcus aureus was lower (MIC 4 mg/mL). Hesperetin (0.165 mg/mL) reduced ability of Candida to form biofilms and moderately reduced exopolysaccharide levels in biofilm matrix. Effect on the eradication of 24 h old C. albicans biofilms was promising at 1.320 mg/mL. Inhibition of staphylococcal biofilm formation required higher concentrations of hesperetin (<50% inhibition with MIC 4 mg/mL). Establishment of polymicrobial C. albicans-S. aureus biofilm was significantly inhibited with the lowest examined hesperetin concentration (1 mg/mL) in crystal violet and CFU assays. Hesperetin toxicity was examined towards MRC-5 fibroblasts (IC50 0.340 mg/mL) and in brine shrimp lethality assay (LC50 > 1 mg/mL). Hesperetin is efficient in combating growth and biofilm formation of Candida species. However, its antibacterial application should be further examined due to the cytotoxic effects provoked in the antibacterial concentrations.

Cinnamon essential oil and its emulsion as efficient antibiofilm agents to combat Acinetobacter baumannii.

Acinetobacter baumannii is an emerging nosocomial pathogen resistant to a wide spectrum of antibiotics, with great potential to form a biofilm, which further aggravates treatment of infections caused by it. Therefore, searching for new potent agents that are efficient against A. baumannii seems to be a necessity. One of them, which has already been proven to possess a wide spectrum of biological activities, including antimicrobial effect, is cinnamon essential oil. Still, further increase of antibacterial efficacy and improvement of bioavailability of cinnamon oil is possible by emulsification process. The aim of this study was comparative analysis of cinnamon essential oil and its emulsion against biofilm forming A. baumannii clinical isolates. Furthermore, the investigation of toxicological aspects of possible applications of essential oil and emulsion was done as well. Gas chromatography-mass spectrometry of essential oil indicated trans-cinnamaldehyde as the most abundant component. The cinnamon emulsion was synthesized from cinnamon essential oil by combining modified low- and high- energy methods. Synthesized emulsion was characterized with Fourier-transform infrared spectroscopy and photon correlation spectroscopy. Both substances exhibited significant antibacterial (minimal inhibitory concentrations in the range 0.125-0.5 mg/ml) and antibiofilm effects (inhibitions of formation and reduction of pre-formed biofilm were 47-81 and 30-62%, respectively). Compared to essential oil, the efficacy of emulsion was even stronger considering the small share of pure oil (20%) in the emulsion. The result of biofilm eradication assay was confirmed by scanning electron microscopy. Even though the cytotoxicity was high especially for the emulsion, genotoxicity was not determined. In conclusion, strong antibacterial/antibiofilm effect against A. baumannii of the cinnamon essential oil and the fact that emulsification even potentiated the activity, seems to be of great significance. Observed cytotoxicity implicated that further analysis is needed in order to clearly determine active principles being responsible for obtained antibacterial/antibiofilm and cytotoxic properties.

Essential Oils of Three Balkan Abies Species: Chemical Profiles, Antimicrobial Activity and Toxicity toward Artemia salina and Drosophila melanogaster.

This study focused on the essential oils (EOs) isolated from needles with twigs of three indigenous Balkan Abies species (A. alba, A. × borisii-regis and A. cephalonica) regarding their chemical composition, antimicrobial activity and toxicity toward crustaceans and insects. Even though distinct phytochemical profiles of dominant volatiles were revealed for each species, ß-pinene and α-pinene represented the first two major volatiles in all three EOs. Antimicrobial activity of EOs has shown inhibitory effect against all 17 studied strains (ATCC and respiratory isolates) in the range of 0.62-20.00 mg/mL (MICs). Further, all three EOs exhibited strong toxicity (LC50 <100 µg/mL) in Artemia salina lethality bioassay, but with significant differences that depended on the EO type. Additionally, tested EOs have shown a certain level of toxicity against Drosophila melanogaster, mostly at the highest tested concentration (3 %) which caused significant prolongation of developmental time, larvicidal effect and pupal mortality. In the three biological assays performed, there was no observed inhibitory effect or weakest activity for A. alba EO. Further, A. cephalonica EO has shown the highest levels of antimicrobial activity and toxicity toward A. salina, while in relation to the insecticidal potential, A. cephalonica and A. × borisii-regis EOs exhibited similar level of toxicity against D. melanogaster.

Tamjanika, a Balkan native variety of Vitis vinifera L.: Chemical characterization, antibacterial, and anti-dermatomycosis potential of seed oil.

This study was designed to explore functional food properties of edible seed oil obtained from Tamjanika seeds-autochthonous grape variety of Balkan Peninsula. In order to accomplish our goals, seed oil was isolated by Soxhlet apparatus and chemically characterized regarding fatty acids, carotenoids, tocopherols, and tocotrienols. Antimicrobial activity of the isolated oil was tested by microdilution method. For that purposes, six bacterial species were used, belonging to human infectious agents and food contaminants. Furthermore, the activity of the oil was investigated against clinical isolates of dermatomycetes. Our study has shown that oil of Vitis vinifera L. Tamjanika variety was an abundant source of polyunsaturated fatty acids (81.43%) with predominant linoleic acid. HPLC analysis revealed the presence of carotenoid lutein (0.15 mg/100 g). The seed oil was rich in tocotrienols (85.04 mg/100 g) predominating over tocopherols (8.37 mg/100 g). The oil possessed microbicidal activity against all the tested microbes. Bacteria were more sensitive to the effect of the oil (minimum inhibitory concentration [MIC] 7.7-15.4) when compared with oil effect on tested dermatomycetes (MIC 20-40). Our investigation has shown for the first time that grape oil could be active against wide spectrum of bacteria and clinically isolated dermatomycetes. The significance of this study lies in the fact that it pointed out the functional food properties of grape seed oil that was fully chemically characterized.

Elucidating the antibiofilm activity of Frangula emodin against Staphylococcus aureus biofilms.

AIMS: Because the Staphylococcus aureus is one of the most well-known pathogens associated with medical devices and nosocomial infections, the aim of the study was to examine antibiofilm potential of emodin against it. METHODS AND RESULTS: Antibacterial activity was examined through microdilution assay. Antibiofilm testing included crystal violet staining of biofilm biomass and morphology analysis by Atomic force microscopy (AFM). Furthermore, aerobic respiration was monitored using the Micro-Oxymax respirometer. For investigation of gene expression qRT-PCR was performed. Emodin demonstrated strong antibacterial activity and ability to inhibit biofilm formation of all tested strains. The effect on preformed biofilms was spotted in few strains. AFM revealed that emodin affects biofilm structure and roughness. Monitoring of respiration under emodin treatment in planktonic and biofilm form revealed that emodin influenced aerobic respiration. Moreover, qRT-PCR showed that emodin modulates expression of icaA, icaD, srrA and srrB genes, as well as RNAIII, and that this activity was strain-specific. CONCLUSION: The results obtained in this study indicate the novel antibiofilm activity of emodin and its multiple pathways of action. SIGNIFICANCE AND IMPACT OF STUDY: This is the first study that examined pathways through which emodin expressed its antibiofilm activity.

Chemical and Biological Investigations of Allium scorodoprasum L. Flower Extracts.

This study was designed to investigate the impact of different extraction solvent systems on the chemical composition and biological activities of Allium scorodoprasum L. (Amaryllidaceae)-the medicinal plant that was traditionally used as a remedy in the medieval period in the Balkans. Targeted chemical analysis of nine different extracts was performed by UHPLC(-)HESI-QqQ-MS/MS. Antimicrobial and antibiofilm activities of the extracts were investigated on sixteen clinical isolates of bacteria, yeasts and dermatomycetes, all isolated from infected human skin and corneal formations. Cytotoxicity and wound-healing properties were tested on human immortalized keratinocytes (HaCaT cell line). Antioxidant activity was assessed by six different assays, while beneficial potential against certain neurodegenerative diseases and type 2 diabetes was determined in selected enzyme inhibition assays coupled with molecular modeling. The results showed that the obtained extracts were rich in phenolic compounds, especially flavonoid glycosides such as rutin and kaempferol 3-O-glucoside. All of the extracts showed antimicrobial, wound-healing, antioxidant and anti-enzymatic properties. This study is the first of its kind, linking the medieval medicinal use of wild-growing flowers of A. scorodoprasum with contemporary in vitro scientific approaches.

Cymbopogon citratus essential oil: an active principle of nanoemulsion against Enterococcus faecalis root canal biofilm.

Aim: The objective was to formulate and characterize the nanoemulsion based on Cymbopogon citratus oil, intended for use in infected teeth root canal therapy. The investigation of the antioxidant and antibiofilm potential toward Enterococcus faecalis was aimed as well. Materials & methods: Characterization of oil (by GC/MS analysis) and nanoemulsion (by dynamic light scattering instrument), and determination of antibacterial (by microdilution assay), antibiofilm (by crystal violet assay) and antioxidant properties (by 2,2-diphenyl-1-picryl-hydrazyl-hydrate and thiobarbituric acid assay methods) were provided. Antibiofilm efficacy of irrigation procedure including nanoemulsion was screened on extracted teeth (by CFU-counting assay). Results: Notable antibacterial and antibiofilm activity, both against forming and preformed biofilms of oil, was observed. Irrigation involved nanoemulsion showed remarkable antibiofilm potential. Both substances induced some antioxidant activity. Conclusion: Results encourage further research with the aim of application of the nanoemulsion in dental practice.

In Vitro Study of Two Edible Polygonoideae Plants: Phenolic Profile, Cytotoxicity, and Modulation of Keap1-Nrf2 Gene Expression.

Polygonum aviculare and Persicaria amphibia (subfam. Polygonoideae) are used in traditional cuisines and folk medicine in various cultures. Previous studies indicated that phytochemicals obtained from Polygonoideae plants could sensitize chemoresistant cancer cells and enhance the efficacy of some cytostatics. Here, the cytotoxic properties of chemically characterized ethanol extracts obtained from P. aviculare and P. amphibia, individually and in combination with doxorubicin (D), were determined against hepatocarcinoma HepG2 cells. Phenolic composition, cell viability, cell cycle, apoptosis, and the expression of Keap1 and Nrf2 were examined by following methods: LC-MS/MS, LC-DAD-MS, MTT, flow cytometry, and qRT-PCR. Extracts were rich in dietary polyphenolics. Synergistic cytotoxicity was detected for extracts combined with D. The observed synergisms are linked to the interference with apoptosis, cell cycle, and expression of Keap1-Nrf2 genes involved in cytoprotection. The combined approach of extracts and D could emerge as a potential pathway of chemotherapy improvement.

Pink oyster mushroom Pleurotus flabellatus mycelium produced by an airlift bioreactor-the evidence of potent in vitro biological activities.

Four types of mycelial extracts were derived from the airlift liquid fermentation (ALF) of Pleurotus flabellatus, namely exopolysaccharide (EX), endopolysaccharide (EN), hot water (WE), and hot alkali (AE) extracts. Such extracts were screened for their active components and biological potential. EN proved to be most effective in inhibition of lipid peroxidation (EC50 = 1.71 ± 0.02 mg/mL) and in Cupric ion reducing antioxidant capacity (CUPRAC) assay (EC50 = 2.91 ± 0.01 mg TE/g). AE exhibited most pronounced ability to chelate ferrous ions (EC50 = 4.96 ± 0.08 mg/mL) and to scavenge ABTS radicals (EC50 = 3.36 ± 0.03 mg TE/g). ß-glucans and total phenols contributed most to the chelating ability and quenching of ABTS radicals. Inhibition of lipid peroxidation correlated best with total glucans, total proteins, and ß-glucans. Total proteins contributed most to CUPRAC antioxidant capacity. Antifungal effect was determined against Candida albicans ATCC 10231 (MIC: 0.019-0.625 mg/mL; MFC: 0.039-2.5 mg/mL), and towards C. albicans clinical isolate (MIC and MFC: 10.0-20.0 mg/mL). Comparison of cytotoxicity against colorectal carcinoma HCT 116 cells (IC50: 1.8 ± 0.3-24.6 ± 4.2 mg/mL) and normal lung MRC-5 fibroblasts (IC50: 17.0 ± 4.2-42.1 ± 6.1 mg/mL) showed that EN, and especially AE possess selective anticancer activity (SI values 3.41 and 9.44, respectively). Slight genotoxicity was observed only for AE and EX, indicating the low risk concerning this feature. Notable antioxidative and anticandidal activities, selective cytotoxicity against colorectal carcinoma cells, and absence/low genotoxicity pointed out that ALF-cultivated P. flabellatus mycelium could be considered as a valuable source of bioactive substances.

New insight into the antigenotoxic activity of Gentiana lutea extracts - Protective effect against food borne mutagens.

Food mutagens formed from amino acids during heating of meat have the potential to induce serious consequences on human health. As a result, the identification of naturally occurring, genoprotective agents, is of great importance. The aim of this study was to chemically characterize a root and leaf extracts of Gentiana lutea and to investigate the antigenotoxic effects of extracts and pure constituents (gentiopicroside and mangiferin). Antigenotoxic effects were shown for combinations with the food borne mutagens IQ and PhIP using hepatoma HepG2 cells. Furthermore, their antioxidant activity and their capacity to modulate Nrf2 expression and affect the glutathione redox status were tested. Chemical analyses showed that the most abundant constituents found in root extract are gentiopicroside and sweroside. On the other hand, homoorientin and isovitexin were the dominant ones in leaf extract. Strong genoprotective activities of all tested compounds against both mutagens were observed in alkaline comet assays (up to 77% of tail intensity inhibition, p < 0.001). The protection against glutathione depletion was partially due to the radical scavenging activity and up-regulation of Nrf2 expression by the substances. The results of this study strongly encourage further investigations of the antimutagenic properties of G. lutea.

Antimicrobial potential of irrigants based on essential oils of Cymbopogon martinii and Thymus zygis towards in vitro multispecies biofilm cultured in ex vivo root canals.

OBJECTIVE: The objective was to determine the antibiofilm efficacy of Cymbopogon martinii and Thymus zygis essential oils and to estimate the disruption potential of oil-based endodontic irrigants on the multispecies biofilm formed in the root canals of extracted teeth. DESIGN: The essential oils were characterized (GC-MS), while their antibacterial and cytotoxic properties were detected by microdilution and MTT assays. Particles sizes and polydispersity indices were determined for the irrigants. The isolates from root canals of pediatric patients were identified (MALDI-TOF). The multispecies biofilms were formed from Streptococcus mitis, Streptococcus sanguinis and Enterococcus faecalis. Antibiofilm properties of the essential oils and irrigants were determined by crystal violet and plate counting assay. Statistical analysis was performed by one-way ANOVA. RESULTS: The screening of biofilm biomass revealed a disruption potential of C. martinii essential oil. The plate counting assay showed the efficacy of both oils in diminishing cell viability: high in biofilms (reduction of log10CFU was 2.75-2.87) and moderate in the planktons formed above. The essential oil-based irrigants showed the same antibiofilm activity as in the control (1.5 % sodium hypochlorite) for C. martini and almost 2-fold higher for T. zygis. Successive irrigations with 1.5 % sodium hypochlorite, saline and an oil-based irrigant was more efficient for C. martini than for the control (reductions of log10CFU was 1.69 and 1.14, respectively). CONCLUSIONS: Notable activities achieved by essential oils and oil-based irrigants, particularly regarding C. martini, against the biofilm consisting of S. mitis, S. sanguinis, and E. faecalis, are expected to encourage further research in endodontics.

Anti-Virulence Potential and In Vivo Toxicity of Persicaria maculosa and Bistorta officinalis Extracts.

Many traditional remedies represent potential candidates for integration with modern medical practice, but credible data on their activities are often scarce. For the first time, the anti-virulence potential and the safety for human use of the ethanol extracts of two medicinal plants, Persicaria maculosa (PEM) and Bistorta officinalis (BIO), have been addressed. Ethanol extracts of both plants exhibited anti-virulence activity against the medically important opportunistic pathogen Pseudomonas aeruginosa. At the subinhibitory concentration of 50 µg/mL, the extracts demonstrated a maximal inhibitory effect (approx. 50%) against biofilm formation, the highest reduction of pyocyanin production (47% for PEM and 59% for BIO) and completely halted the swarming motility of P. aeruginosa. Both extracts demonstrated better anti-quorum sensing and antibiofilm activities, and a better ability to interfere with LasR receptor, than the tested dominant extracts' constituents. The bioactive concentrations of the extracts were not toxic in the zebrafish model system. This study represents an initial step towards the integration of P. maculosa and B. officinalis for use in the treatment of Pseudomonas infections.

Discriminant analysis of cardiovascular and respiratory variables for classification of road cyclists by specialty.

BACKGROUND: Different variables determine the performance of cyclists, which brings up the question how these parameters may help in their classification by specialty. The aim of the study was to determine differences in cardiorespiratory parameters of male cyclists according to their specialty: flat riders (N.=21), hill riders (N.=35), or sprinters (N.=20) and obtain the multivariate model for further cyclists classification by specialties, based on selected variables. METHODS: Seventeen variables were measured at submaximal and maximum load on the cycle ergometer Cosmed E 400HK (Cosmed, Rome, Italy) (initial 100 W with 25-W increase, 90-100 rpm). Multivariate discriminant analysis was used to determine which variables group cyclists within their specialty, and to predict which variables can direct cyclists to a particular specialty. RESULTS: Among nine variables that statistically contribute to the discriminant power of the model, achieved power on the anaerobic threshold and the produced CO2 had the biggest impact. The obtained discriminatory model correctly classified 91.43% of flat riders, 85.71% of hill riders, while sprinters were classified completely correct (100%), i.e. 92.10% of examinees were correctly classified, which point out the strength of the discriminatory model. CONCLUSIONS: Respiratory indicators mostly contribute to the discriminant power of the model, which may significantly contribute to training practice and laboratory tests in future.

Beneficial Effect of Virgin Coconut Oil on Alloxan-Induced Diabetes and Microbiota Composition in Rats.

The gut microbiota is composed of a diverse population of obligate and facultative anaerobic microorganisms which are shown to influence host metabolism and immune homeostasis. This study investigated the effects of virgin coconut oil on the weekly fasting glycaemia, daily food and water intake and weekly body mass gain over 16 weeks, as well as the changes in composition of gut microbiota in both non-diabetic and alloxan-induced diabetic rats. Although the intake of virgin coconut oil did not decrease the diabetes-induced hyperglycemia, it affected the secondary parameters, such as food and water intake and average body mass gain. Furthermore, its potential to positively affect the fecal microbiome was proved, since it significantly increased the abundance of probiotic bacteria, such as Lactobacillus, Allobaculum and Bifidobacterium species.