Synergistic interactions of essential oil components with antibiotics against multidrug-resistant Corynebacterium striatum.

AIM: In this study, it was aimed to examine the antibacterial activity of the essential oil components (EOCs), carvacrol (CAR), cinnamaldehyde (CIN), thymol (TH), alpha pinene (α-PN), eucalyptol (EU), limonene (LIM), and the antibiotics, linezolid (LZD), vancomycin (VAN), gentamicin (GEN), ciprofloxacin (CIP), clindamycin (CLN), and penicillin (PEN) against 50 multidrug resistant Corynebacterium striatum strains, and the synergistic interactions of CAR and CIN with the antibiotics against 10 randomly selected Coryne. striatum strains to explore synergistic interactions to determine if their combined use could enhance antibiotic activity and potentially reduce resistance. METHODS AND RESULTS: The activity of the EOCs and the antibiotics against Coryne. striatum strains isolated from clinical specimens, was examined by broth microdilution method. The synergistic interactions of the EOCs with the antibiotics against 10 randomly selected Coryne. striatum strains were determined by checkerboard method. EOCs, CIN, and CAR and antibiotics, LZD, VAN, GEN, CIP, and CLN were detected to have antibacterial activity against Coryne. striatum strains alone and either synergistic interactions were observed in combinations of the antibiotics with EOCs. CONCLUSIONS: All Coryne. striatum strains were determined to be susceptible to VAN and LZD and resistant to GEN, PEN, CIP, and CLN. Synergistic interactions were observed in all combinations of antibiotics tested with CAR and CIN.

[Comparison of Conventional Methods with Molecular Methods in the Diagnosis of Trichomonas vaginalis and Investigation of Metronidazole Resistance]. / Trichomonas vaginalis'in Tanisinda Konvansiyonel Yöntemlerin Moleküler Yöntemlerle Karsilastirilmasi ve Metronidazol Direncinin Arastirilmasi.

Trichomoniasis is a sexually transmitted parasitic infection caused by Trichomonas vaginalis. In the diagnosis of trichomoniasis, direct microscopy (DM) is preferred, which is a cheap and fast method, although it has low sensitivity. Culture methods, which are accepted as the gold standard, can only be applied in certain centers due to the need for experienced personnel and the ability to get results within 2-7 days, despite their high sensitivity. In this study, it was aimed to compare conventional microscopic and culture methods used in the routine diagnosis of T.vaginalis with polymerase chain reaction (PCR) method and to investigate ntr4 and/or ntr6 gene polymorphism in the nitroreductase gene region, which are thought to be associated with metronidazole resistance in T.vaginalis strains isolated from clinical specimens. Vaginal swab specimens were collected from the posterior fornix of the vagina with two sterile ecuvion sticks during the gynecological examinations of 200 patients who applied to the Balikesir University Health Practice and Research Hospital, Obstetrics and Gynecology Polyclinic between March 2019 and August 2021. The first swab sample was used for direct microscopic examination, Giemsa staining and conventional PCR analysis, while the second swab specimen was taken into trypticase-yeast-extract-maltose (TYM) medium for T.vaginalis culture and followed for eight days at 37 °C. All specimens were screened for the presence of T.vaginalis using primers specific to the ß-tubulin (btub1) gene region and clinical isolates grown in TYM medium were examined for metronidazole resistance using primers specific for the nitroreductase gene region by using conventional PCR. Drug resistance test was also performed for the isolates in which polymorphism associated with metronidazole resistance was detected. Eight (4%) of 200 patient specimens were found positive by both culture/staining and PCR methods. The mean age of the patients included in the study was 39.9, while the mean age of the patients with positive T.vaginalis was 41.8. The most common clinical findings in the patients were foul-smelling vaginal discharge (36%), groin pain (21%), vaginal itching (19%), and burning sensation during urination (18%). In three out of eight T.vaginalis strains isolated from clinical samples, the presence of polymorphism in the ntr6 gene, which is thought to be associated with metronidazole resistance, was demonstrated by PCR. It was observed that three isolates with ntr6 gene polymorphism were phenotypically resistant to metronidazole (MLK= 390 µM). In this study, the fact that three of eight clinical isolates that were resistant to metronidazole by the broth microdilution method and as well as showing ntr6 gene polymorphism supported the thesis that there might be a close relationship between metronidazole resistance and ntr6 gene polymorphism. As a result, the use of culture and molecular methods in the diagnosis of T.vaginalis, in addition to the microscopy method, may contribute to a more accurate laboratory diagnosis of the agent, to detect metronidazole resistance molecularly and phenotypically, to determine metronidazole resistance rates in our country and to update treatment protocols within the framework of these data.

[Antibacterial Activity and Synergistic Interaction of Various Essential Oil Components and Antibiotics]. / Çesitli Uçucu Yag Bilesenleri ile Antibiyotiklerin Antibakteriyel Etkinligi ve Sinerjik Etkilesimi.

Inappropriate and uncontrolled use of antibiotics in humans and animals leads to the emergence of multi-resistant bacteria. Before the discovery of antibiotics, plant extracts and essential oils were used for therapeutic purposes. Today, due to increasing antibiotic resistance, many studies are frequently carried out on the antimicrobial activities of natural active substances that can be a source for new drug candidates. The aim of this study was to investigate the antibacterial activity of components such as α-pinene (α-PN), p-cymene (p-CYM), carvacrol (CAR), thymol (TY) and eugenol (EG) found in the essential oils of many plants and their synergistic interaction with antibiotics. In this study, the antibacterial activity of these essential oil components and antibiotics in clinical use such as gentamicin (GEN), tetracycline (TET), tigecycline (TGC) and linezolid (LZD), against Staphylococcus aureus [methicillin resistant S.aureus (MRSA), and methicillin sensitive S.aureus (MSSA)], Escherichia coli and Pseudomonas aeruginosa strains were determined by disc diffusion and microdilution method. In addition, the interaction between the essential oil components and antibiotics was also determined by the checkerboard method. While CAR, TY and EG components showed strong antibacterial activity, the antibacterial activity of αPN and p-CYM was found to be weak. Combinations of α-pinene, carvacrol, thymol and eugenol with gentamicin and tetracycline mostly showed synergistic interactions against all bacteria. In αPN, CAR, TY and EG with GEN and TET, synergistic/partial synergistic interaction was observed against S.aureus strains, while indifferent interaction was detected in E.coli and P.aeruginosa strains. The combination of αPN and p-CYM with TGC showed synergistic interaction against E.coli and P.aeruginosa strains, and additive and indifferent interaction against S.aureus strains. On the other hand, synergistic interaction was observed against all bacterial strains in combinations of TGC and CAR, TY and EG components. Antagonistic interaction was not detected in any of the tested component-antibiotic combinations against the bacteria used in our study. A synergistic interaction between natural bioactive components and commonly used antibiotics may contribute to the effectiveness of antibiotics and components at lower doses, minimizing their potential toxic side effects and reducing treatment costs. However, more research is needed in terms of their pharmacokinetic and toxic properties to evaluate the therapeutic application potential of phytochemicals.

Evaluation of in vitro activity of carvacrol against Candida albicans strains.

In this study, the in vitro activity of carvacrol on 100 clinical isolates of Candida albicans was evaluated by determining the minimum inhibitory concentrations (MICs). Susceptibility of clinical isolates of C. albicans to carvacrol was determined by broth microdilution method. Carvacrol was active in vitro against all strains of C. albicans, with MICs ranging from 0.125% to 0.004%. MIC(50) and MIC(90) values of carvacrol were observed at 0.064 and 0.125 mg mL(-1), respectively. MIC and MFC values were similar for most of the tested strains. These results indicated that carvacrol inhibited the growth of C. albicans, implying its potential use in the topical treatment of C. albicans infections.

Chemical composition, antibacterial and antifungal activity of the essential oil of Thymbra spicata L. from Turkey.

The composition of the essential oil from aerial parts of Thymbra spicata L. from Turkey was analysed by GC-MS and its in vitro antimicrobial activity was examined. GC-MS analysis of the essential oil resulted in the identification of twenty-three constituents, representing 97.04% of the oil. The major compounds detected in the essential oil were carvacrol (60.39%), gamma-terpinene (12.95%), and p-cymene (9.61%). The in vitro efficacy of the essential oil against 21 bacteria and seven Candida species was examined using disc diffusion and minimum inhibitory concentration (MIC) methods. The essential oil demonstrated strong anti-microbial activity in a wide spectrum against most microorganisms, particularly the yeasts tested. This is the first report on the anticandidal properties of the essential oil of T. spicata. In conclusion, this study confirms that T. spicata essential oil could be considered as a natural antimicrobial source.

Antibacterial activity and phytochemical evidence for the plant origin of Turkish propolis from different regions.

Honeybees collect propolis from practically any abundant plant source in the neighborhood of the hive, be it populus, eucalyptus, pine, sugarcane, cashew nut or orange trees. We have described that the origin plants of Turkish propolis are Populus sp., Eucalyptus sp. and Castanea sativa. In our previous study, propolis samples from Middle Anatolia displayed the typical pattern of "poplar" propolis: they contained pinobanksin, caffeic and ferulic acids and their esters. The propolis samples examined in this study were shown not to contain polar phenolics. The main components of Eucalyptus propolis were aromatic acids, mainly cinnamic acid and its esters, that are usually found in Eucalyptus species resins. The second distinct sample originated from West Anatolia. Although it contained low amounts of phenolic substances and aromatic acids, its main components were sugars and glycosides. The study revealed that there was no significant difference between propolis samples in antibacterial activity, however the yeasts were shown to be more sensitive to eucalyptus-propolis. Gram negative bacteria were susceptible to none of the samples tested.

Antimicrobial activity of essential oil and methanol extracts of Achillea sintenisii Hub. Mor. (Asteraceae).

The essential oil, obtained by Clevenger distillation, and water-soluble and water-insoluble parts of the methanol extracts of Achillea sintenisii Hub. Mor. were individually assayed for their antimicrobial activities against 12 bacteria and two yeasts, Candida albicans and C. krusei. No activity was exhibited by the water-soluble subfraction, whereas both the water-insoluble subfraction of the methanol extracts and the essential oil were found to be active against some test microorganisms studied. Since the essential oil possessed stronger activity than the other extracts tested, it was further fractionated and the fractions were evaluated for their antimicrobial activity, followed by GC-MS analysis, resulting in the identification of 32 compounds which constituted 90.2% of the total oil. The GC-MS analysis of the oil and its fractions revealed that the main components of the oil, e.g. camphor and eucalyptol, possessed appreciable activity against C. albicans and Clostridium perfringens. The fi ndings presented here also suggest that the other constituents of the oil, e.g. borneol and piperitone can also be taken into account for the activity observed.

Antimicrobial and antioxidant activity of the essential oil and methanol extracts of Thymus pectinatus Fisch. et Mey. Var. pectinatus (Lamiaceae).

The essential oil, obtained by using a Clevenger distillation apparatus, and water-soluble (polar) and water-insoluble (nonpolar) subfractions of the methanol extract of Thymus pectinatus Fisch. et Mey. var. pectinatus were assayed for their antimicrobial and antioxidant properties. No (or slight) antimicrobial activity was observed when the subfractions were tested, whereas the essential oil showed strong antimicrobial activity against all microorganisms tested. Antioxidant activities of the polar subfraction and the essential oil were evaluated using 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical, superoxide radical scavenging, and lipid peroxidation assays. The essential oil, in particular, and the polar subfraction of the methanol extract showed antioxidant activity. The essential oil was analyzed by GC/MS, and 24 compounds, representing 99.6% of the essential oil, were identified: thymol, gamma-terpinene, p-cymene, carvacrol, and borneol were the main components. An antimicrobial activity test carried out with fractions of the essential oil showed that the activity was mainly observed in those fractions containing thymol, in particular, and carvacrol. The activity was, therefore, attributed to the presence of these compounds. Other constituents of the essential oil, such as borneol, gamma-terpinene, and p-cymene, could be also taken into account for their possible synergistic or antagonistic effects. On the other hand, thymol and carvacrol were individually found to possess weaker antioxidant activity than the crude oil itself, indicating that other constituents of the essential oil may contribute to the antioxidant activity observed. In conclusion, the results presented here show that T. pectinatus essential oil could be considered as a natural antimicrobial and antioxidant source.