Comparison of the Intensity of Biofilm Production by Oral Microflora and Its Adhesion on the Surface of Zirconia Produced in Additive and Subtractive Technology: An In Vitro Study.

BACKGROUND: This in vitro study set out to find out how well oral cavity-dwelling bacteria can form biofilms and adhere on the surfaces of zirconium oxide samples created by 3D printing and milling technologies. METHODS: 5 strains of microorganisms were used for the study, and 40 zirconium oxide samples were prepared, which were divided into two groups (n = 20)-20 samples produced using removal technology comprised the control group, while 20 samples produced by 3D printing technology comprised the test group. The prepared samples were placed in culture media of bacteria and fungi that naturally occur in the oral cavity. Then, the intensity of biofilm build-up on the samples was determined using qualitative and quantitative methods. The results for both materials were compared with each other. RESULTS: No variations in the degree of biofilm deposition on zirconium oxide samples were found for the microorganisms Streptococcus mutans, Pseudomonas aeruginosa, Enterococcus faecalis, and Staphylococcus aureus. For Candida albicans fungi, more intense biofilm deposition was observed on samples made using 3D printing technology, but these differences were not statistically significant. CONCLUSION: The biofilm accumulation intensity of ceramics produced by additive technology is comparable to that of milled zirconium oxide, which supports the material's broader use in clinical practice from a microbiological perspective. This ceramic has demonstrated its ability to compete with zirconium oxide produced by milling techniques in in vitro experiments, but sadly, no in vivo tests have yet been found to determine how this material will function in a patient's oral cavity.

Similarity Analysis of Klebsiella pneumoniae Producing Carbapenemases Isolated from UTI and Other Infections.

Klebsiella pneumoniae is an important opportunistic pathogen responsible for severe infections, mainly urinary tract infections (UTIs) and pneumonia. Hospital epidemic infections caused by multiresistant strains of carbapenemase-producing K. pneumoniae are the most concerning. NDM-producing strains are resistant to a wide range of antibiotics and have become the most significant threat. Determining the natural reservoirs and routes of infections is essential to end hospital outbreaks. Understanding the relatedness of K. pneumoniae strains is essential to determine the range and nature of the infection. The study compared phylogenetic relatedness between multiresistant K. pneumoniae strains isolated from hospitalized patients. Susceptibility to drugs and mechanisms of resistance were confirmed using phenotypic methods. PFGE was used to analyze the relatedness between strains. We analyzed 69 K. pneumoniae strains from various healthcare units. The isolates were mainly identified from urine. Strains were resistant to ß-lactam antibiotics with ß-lactamase inhibitors, cephalosporins, and quinolones. Their susceptibility to aminoglycosides and carbapenem antibiotics was diverse. Most of the isolated strains produced New Delhi metallo-ß-lactamase (NDM). Although K. pneumoniae strains were classified into several genotype clusters, closely related isolates were confirmed in the same hospital's wards, and in two hospitals in the same province.

Enhancement of neutrophil chemotaxis by trans-anethole-treated Staphylococcus aureus strains.

This study aimed to analyze the chemotactic response of differentiated HL-60 neutrophil-like (dHL-60) cells to trans-anethole (TA)-treated Staphylococcus aureus strains. Special attention was paid to evaluate the influence of TA on the chp gene expression level, as well as molecular docking and molecular dynamics (MD) simulation studies on interactions of TA with chemotaxis inhibitory protein of S. aureus (CHIPS). The following parameters were studied: susceptibility to TA using the agar diffusion method, the chp gene detection and its expression under TA influence, and clonal diversity of S. aureus strains using molecular techniques. Furthermore, a chemotactic response of dHL-60 cells to TA-treated S. aureus using Boyden chamber assay was detected and molecular modeling using both the docking methodology and unbiased MD simulations was conducted. It was found that TA showed antibacterial activity against all strains. Three genotypes and one unique pattern were distinguished among the strains. 50% of the isolates were chp-positive. It was observed that TA reduced/inhibited chp gene expression in most S. aureus strains. Enhanced chemotactic response of dHL-60 cells to TA-treated S. aureus strains was also noted. This correlation was similar for both chp-positive and chp-negative strains. Both molecular docking and MD simulations studies confirmed that TA is preferentially bound in the complement component 5a/CHIPS interface interaction region and can interfere with any processes exploiting this binding cavity. It has been proven that dHL-60 cells exhibited a higher chemotactic response to TA-treated S. aureus strains in comparison to non-treated bacteria, regardless of the achieved expression of the chp gene or its lack. Nevertheless, further analyses are required to understand this mechanism better.

Immunomodulatory effects of trans-anethole-treated Staphylococcus aureus Newman strain.

In our former studies based on a human whole-blood model infected with trans-anethole (TA)-treated Staphylococcus aureus Newman strain, we have observed that selected parameters/mechanisms of innate and acquired immune response were more enhanced in comparison to samples infected with non-treated bacteria. Due to this observation, the current study aimed to evaluate the concentration of selected proteins involved in both types of responses (IL-1α, IL-1ß, IL-2, IL-6, IL-12, IL-17, TNF-α, IFN-γ, G-CSF, C5a, CCL1-CCL5, CXCL1, CXCL2, CXCL9-CXCL11, MMP-8, TLR2, and PGLYRP1) in healthy participants' plasma after blood stimulation of TA-treated S. aureus Newman strain. Determination of analyzed protein concentration was conducted using Luminex and ELISA assays. Based on the results, it has been proven that the immunomodulatory potential of TA-treated S. aureus Newman strain on increasing IL-1ß, IL-6, TNF-α, IL-12, G-CSF, C5a, CCL2-CCL4, CXCL1, CXCL2, MMP-8 and PGLYRP1 levels in plasma. Moreover, it has been also demonstrated an association between TNF-α and CCL4 in a blood model infected with TA-treated cells. More research is warranted to find more underlying mechanisms involved in the effects of TA-treated S. aureus Newman in human blood, mainly whether the observed "immunity boost" can be regulated after bacteria elimination. Therefore, the potential of TA should be further explored to understand under which conditions it might help treat or prevent infections caused by S. aureus.

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.

Staphyloxanthin inhibitory potential of trans-anethole: A preliminary study.

The reduction of staphyloxanthin (STX) production in Staphylococcus aureus under trans-anethole (TA) influence was proven in former studies. However, no tests concerning the impact of TA on a biosynthetic pathway of this carotenoid pigment have been published so far. Thus, for the first time, the present preliminary study evaluated the influence of TA on the expression level of genes (crtOPQMN operon and aldH) encoding STX pathway enzymes. Additional attention was paid to the identification of STX and its intermediates. Gene expression and identification of extracted compounds were conducted using quantitative real-time PCR and HPLC-MS techniques, respectively. The analyzes showed no difference in crtM, crtN, crtO, crtP, crtQ, and aldH gene expression between bacterial samples isolated from the non-stimulated (control) medium and the stimulated one with TA. Compared to the control group that showed the presence of all metabolic intermediates and STX, the TA-treated bacteria were characterized by a lack or a significant reduction of the majority of compounds, except 4,4'-diaponeurosporenoate, the content of which was elevated in the TA-treated sample. Moreover, in silico molecular docking analysis revealed that TA is capable to create relatively strong interactions with both 4,4'-diapophytoene synthase and 4,4'-diapophytoene desaturase. The preliminary findings indicate that the previously observed TA effect reducing the number of S. aureus colonies pigmentation is probably not associated with the expression levels of genes encoding STX pathway enzymes. It has been proven that adding TA to the medium can interfere with the formation of STX at different levels of its biosynthetic pathway.

Analysis of the prevalence of colistin resistance among clinical strains of Klebsiella pneumoniae.

INTRODUCTION AND OBJECTIVE: Klebsiella pneumoniae is an essential component of the human gut microflora. However, it can pose a threat by causing opportunistic infections, especially in hospitalised or immunocompromised patients. It is a serious problem for health medicine, primarily because of increasing resistance to previously used antibiotics. Infections with multidrug-resistant strains are difficult to treat, creating a challenge for clinicians. Also of growing concern is the increasing resistance to the drug of last resort - colistin (CL). The aim of the study is to determine the prevalence of resistance to CL among clinical K. pneumoniae strains. MATERIAL AND METHODS: The study was conducted on 200 clinical strains of K. pneumoniae. Drug susceptibility, production of resistance mechanisms, and determination of the minimum inhibitory concentration of CL were evaluated. RESULTS: Of all isolates, 73.0% produced carbapenemases, while the remainder produced an extended substrate spectrum - ß-lactamases (ESBLs). All strains showed a diverse antibiotic resistance profile. Resistance to CL was noted among 14.5% of carbapenemase-producing strains, particularly MBL and OXA-48. ESBL-positive strains showed full susceptibility to CL. CONCLUSIONS: Although a low rate of CL resistance was observed, this was true for strains simultaneously producing carbapenemases. Such strains should be under special epidemiological surveillance due to their potential to cause epidemic outbreaks. Monitoring the prevalence of clinical CL-resistant strains would allow for more effective counteraction against pathogens in various fields, including medicine, agriculture, veterinary medicine and industry.

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.

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.

Antibacterial and Anti-Biofilm Activities of Essential Oil Compounds against New Delhi Metallo-ß-Lactamase-1-Producing Uropathogenic Klebsiella pneumoniae Strains.

The World Health Organization points out that the opportunistic pathogen Klebsiella pneumoniae that causes various infections among others, urinary tract infections (UTIs), is one of the high-priority species due to a global problem of antimicrobial resistance. The aim of this study was to investigate antibacterial and anti-biofilm activities of chosen constituents of essential oils against NDM-1-producing, uropathogenic K. pneumoniae strains. The genes encoding lipopolysaccharide (uge, wabG), adhesin gene fimH (type I fimbriae) and gene encoding carbapenemase (blaNDM-1) for all tested strains were detected by PCR amplification. The K. pneumoniae ATCC BAA-2473 reference strain was uge- and blaNDM-1-positive. The effectiveness of fifteen essential oil compounds (EOCs) (linalool, ß-citronellol, linalyl acetate, menthone, (-)-menthol, (+)-menthol, geraniol, eugenol, thymol, trans-anethole, farnesol, ß-caryophyllene, (R)-(+)-limonene, 1,8-cineole, and carvacrol) was assessed by determining the MIC, MBC, MBC/MIC ratio against K. pneumoniae strains by the microdilution method. Anti-biofilm properties of these compounds were also investigated. Thymol, carvacrol and geraniol exhibited the best antibacterial and anti-biofilm activities against uropathogenic NDM-1-producing K. pneumoniae isolates. Results of our investigations provide a basis for more detailed studies of these phytochemicals on their application against uropathogenic K. pneumoniae.

Epidemiological Analysis of Extended-Spectrum ß-Lactamase-Producing Klebsiella pneumoniae Outbreak in a Neonatal Clinic in Poland.

Klebsiella pneumoniae is one of the most common etiological agents isolated from epidemic outbreaks in neonatal wards. We describe how an extended-spectrum ß-lactamase-producing K. pneumoniae (ESBL-KP) outbreak in a neonatal ward was extinguished. During the outbreak, which lasted over two months, 26 neonates were tested for K. pneumoniae, and 42 environmental swabs were taken. Drug susceptibility was determined for the isolated strains, and their virulence and phylogenetic similarity were checked. ESBL-KP colonization was confirmed in 18 neonates, and six were also confirmed to be infected. All strains isolated from patients represented one clonal type, K. pneumoniae. One strain isolated from an environmental source was determined to be a unique pulsed-field gel electrophoresis pattern. Gestational age and Apgar score were assessed as statistically significant for neonates with ESBL-KP infection. The epidemiological measures taken have been successful, and no further cases appeared. Immediate tightening of hospital hygiene rules, screening of all hospitalized neonates, and cohorting ESBL-KP-positive patients proved effective in controlling and ending the outbreak. The lack of ESBL-KP in the environment suggests that the outbreak was transmitted by colonized hospital staff. This theory could be confirmed by introducing mandatory screening for medical personnel.

The Impact of Intraspecies Variability on Growth Rate and Cellular Metabolic Activity of Bacteria Exposed to Rotating Magnetic Field.

Majority of research on the influence of magnetic fields on microorganisms has been carried out with the use of different species or different groups of microorganisms, but not with the use of different strains belonging to one species. The purpose of the present study was to assess the effect of rotating magnetic fields (RMF) of 5 and 50 Hz on the growth and cellular metabolic activity of eight species of bacteria: Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Enterococcus faecalis, Enterobacter cloacae, Moraxella catarrhalis, and Bacillus cereus. However, contrary to the research conducted so far, each species was represented by at least four different strains. Moreover, an additional group of S. aureus belonging to a single clonal type but representing different biotypes was also included in the experiment. The results showed a varied influence of RMF on growth dynamics and cellular metabolic activity, diversified to the greatest extent in dependence on the bacterial strain exposed to the RMF and to a lesser extent in dependence on the frequency of the generated magnetic field. It was found that, with regard to the exposed strain of the same species, the effect exerted by the RMF may be positive (i.e., manifests as the increase in the growth rate or/and cellular metabolic activity) or negative (i.e., manifests as a reduction of both aforementioned features) or none. Even when one clonal type of S. aureus was used, the results of RMF exposure also varied (although the degree of differentiation was lower than for strains representing different clones). Therefore, the research has proven that, apart from the previously described factors related primarily to the physical parameters of the magnetic field, one of the key parameters affecting the final result of its influence is the bacterial intraspecies variability.

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.

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.

Synergistic effect of fennel essential oil and hydrogen peroxide on bacterial biofilm.

INTRODUCTION: Staphylococcal biofilm formation significantly challenges wound management. The causes of difficult-to-treat wounds are not only methicillin-resistant staphylococci, but also methicillin-sensitive strains with different patterns of resistance. Bacterial biofilm significantly limits the access and activity of antimicrobials used in dermatological infections. AIM: To evaluate the synergistic effect of fennel essential oil (FEO) and H2O2 on biofilm formation by Staphylococcus aureus (MSSA and MRSA) reference strains. MATERIAL AND METHODS: Minimum inhibitory concentration (MIC) values were determined for FEO and H2O2 against S. aureus reference strains by the broth microdilution method. The combined effects of the FEO and H2O2 were calculated and expressed in terms of a fractional inhibitory concentration index (FICI) using the checkerboard method. The FEO composition was analyzed by the GC-MS method. The data were analysed by one-way ANOVA. RESULTS: Decreased MIC values for FEO combined with H2O2 were observed in comparison to FEO itself. The combinations of FEO and H2O2 determined synergistic effects on all S. aureus reference strains. Subinhibitory concentration of FEO alone and in combination with 0.5 MIC of H2O2 significantly decreased the production of biofilm biomass in S. aureus strains and reduced the metabolic activity of attached cells. CONCLUSIONS: Combination of fennel essential oil containing nearly 80% trans-anethole and H2O2 represents a potential for further basic and applied research on wound management.

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 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 effect of fennel essential oil and trans-anethole on antibacterial activity of mupirocin against Staphylococcus aureus isolated from asymptomatic carriers.

INTRODUCTION: Staphylococcus aureus constitutes the most pathogenic species within the staphylococcal genus. Humans remain the major reservoirs of this pathogen which colonizes mostly anterior nares of healthy individuals. AIM: To investigate the effect of fennel essential oil (FEO) and trans-anethole (tA) on antibacterial activity of mupirocin (MUP) against S. aureus strains isolated from asymptomatic carriers. MATERIAL AND METHODS: The content of the FEO was analysed with use of the GC-MS method. The research done on 43 S. aureus isolates with different resistance patterns, obtained from nasal vestibule. Antibacterial activity of MUP in combination with FEO or tA was examined using the agar dilution method and E-test method. The data analysis was done with the Pearson's χ2 test. RESULTS: The chemical composition of FEO was consistent with the European Pharmacopoeia (EP) for the main constituent - tA (77.9%) according to the EP recommendations. Macrolide-lincosamide-streptogramin B resistance phenotype was prevalent among 39.5% of S. aureus isolates. FEO concentrations of 2.0% and 2.5% revealed antibacterial activity against 76.7% of isolates, whereas tA inhibited S. aureus growth at concentrations > 4.0%. The MIC values for MUP combined with FEO as well as for MUP combined with tA were < 0.064 µg/ml for 79.1% and 86.0% of S. aureus isolates, respectively. CONCLUSIONS: Our experiment revealed FEO and tA influence on MUP effectiveness. The combination of MUP with FEO as well as MUP with tA are worth considering to implement in S. aureus eradication procedures. These findings will be useful in designing efficient antistaphylococcal agents which can limit the emergence of antibiotic resistance.

The Effect of Subinhibitory Concentrations of trans-Anethole on Antibacterial and Antibiofilm Activity of Mupirocin Against Mupirocin-Resistant Staphylococcus aureus Strains.

Aim: The aim of this study was to evaluate the effect of subinhibitory concentrations of trans-anethole on antibacterial and antibiofilm properties of mupirocin against mupirocin-resistant Staphylococcus aureus strains. Methods: Following parameters were examined: isolates susceptibility to antibiotics, minimum inhibitory concentration (MIC) of trans-anethole, antibacterial activity of mupirocin/trans-anethole combination, detection of ileS2 gene, genotypic relativity of isolates using pulsed-field gel electrophoresis method, and the influence of mupirocin/trans-anethole combination on S. aureus biofilm formation. Results: Our study revealed that trans-anethole combined with mupirocin increased the growth inhibition zone diameter around the mupirocin disk, independently on S. aureus strains susceptibility to this antibiotic. Moreover, combination of subinhibitory (MIC50) concentration of mupirocin and trans-anethole significantly decreased biofilm biomass. Conclusions: trans-Anethole appeared efficient in increasing susceptibility to mupirocin and decreasing biofilm formation in S. aureus strains used in this study. Reduction of biofilm formation can potentially protect against S. aureus recolonization. Moreover, use of trans-anethole in combination with mupirocin can increase the mupirocin activity against methicillin-resistant and mupirocin-resistant S. aureus strains.