A selection of structurally diverse components of essential oils was subjected to an antibacterial screening against the Grampositive bacteria Staphylococcus aureus and Streptococcus pneumoniae as well as the Gram-negative bacteria Escherichia coli and Haemophilus influenzae using a broth microdilution method. In addition, the irritation potential of these substances to mucous membranes was evaluated by a modified HET-CAM (Hen's Egg Test-Chorioallantoic Membrane) assay. Minimum inhibitory concentrations (MIC) and irritation threshold concentrations (ITC) were compared in order to identify substances with a low mucous membrane irritation potential at the antibacterially effective concentration. Regarding the antibacterial activity, the most active substances were cinnamaldehyde, phenols, monoterpene alcohols and aldehydes. Oxygenated monoterpenes and phenylpropanoids without either alcohol or aldehyde function (ethers, ketones) showed moderate activity. Mono-and sesquiterpene hydrocarbons, as well as sesquiterpene alcohols, were more or less inactive against most of the bacteria tested, with the exception of Streptococcus pneumoniae whose growth was inhibited by sesquiterpenes at exceptionally low concentrations. In most cases, the irritation potential of the test compounds could also be traced back to their structural features. 2-Phenylethanol, monoterpene alcohols, phenols and aromatic aldehydes had the lowest ITC-values, followed by monoterpene aldehydes, ketones and ethers. In contrast, monoterpene hydrocarbons caused irritation only at higher doses. Of special interest was the fact that all the sesquiterpenes tested were not irritant even when applied undiluted to the chorioallantoic membrane (CAM).
The aim of the present study was to investigate the susceptibility of bacteria that play a role in respiratory tract and skin infections to the essential oils of catnip (Nepeta cataria), lemon catnip (N. cataria var. citriodora) and lemon balm (Melissa officinalis) with regard to their chemical composition. In addition, we wanted to assess whether antibiotic-resistant and-sensitive strains differ in their susceptibility to the oils and if there are cross resistances between standard antibiotics and essential oils. To evaluate the safety of topical application, cytotoxicity of the oils was studied in human keratinocyte and bronchial epithelial cell lines and irritation threshold concentrations were determined in ovo using the HET-CAM-test. The composition of the essential oils was analyzed by GC and GC-MS. Their MICs and MBCs were determined by a broth microdilution method against both reference strains from culture collections and clinical isolates with different susceptibility to standard antibiotics. Cytotoxicity was assessed by the MTT assay. Except for P. aeruginosa (MIC ≥2 %), all reference strains tested were susceptible to catnip and lemon balm oils with MIC values ranging from 0.016 % to 0.25 % (v/v). The clinical isolates were as susceptible to the oils (± 1 serial dilution) as the corresponding reference strains, regardless of their origin and resistance to standard antibiotics. The oils were cytotoxic to both keratinocytes and bronchial epithelial cells at CC 50 values from 0.0012% to 0.015% (v/v). Lemon balm oil, whose main components were monoterpene aldehydes, exhibited the highest antibacterial and cytotoxic activity, followed by lemon catnip oil, which contained mainly monoterpene alcohols, and catnip oil, which was characterized by nepetalactones. Our results provide a rationale for the use of catnip, lemon catnip and lemon balm oils in the complementary topical treatment of respiratory tract infections, as the oils show a high antibacterial activity against respiratory tract pathogens, including clinical isolates with reduced susceptibility to standard antibiotics. However, cytotoxicity must be considered in topical therapy.
Brackenridgea zanguebarica is a small tree that is used in traditional African medicine as a type of cure-all for many diseases, including the treatment of wounds. The yellow bark of B. zanguebarica was used for the preparation of an ethanolic extract, which was tested in various concentrations against eleven bacteria, Herpes simplex virus type 1 (HSV-1) and different human tumour cell lines. The extract that contains different polyphenolic substances like calodenin B. Cell growth inhibition, assessed via MTT-assay, was found in all tested human cell lines with IC50 values (concentration of extract that reduced cell viability by 50%) between 33 microg dry extract/mL for HL-60 human myeloid leukaemia cells and 93 microg dry extract/mL for HaCaT human keratinocytes. Staining with Annexin-V-FLUOS and JC-1 followed by subsequent analysis via flow cytometry revealed significant apoptosis-inducing properties. Analysis of caspase activity using a fluorogenic caspase-3 substrate showed a significant caspase activity in Jurkat T-cells after incubation with the extract. The bark extract had a pronounced activity against free HSV-1 and a strong antibacterial activity against Gram-positive strains (MICs: 6-24 microg dry extract/mL), which are often involved in skin infections. Additionally, no irritating properties of the extract could be observed in hen-egg test chorioallantoic membrane (HET-CAM) assay. These findings give a rationale for the traditional use of B. zanguebarica and are a basis for further analysis of the plant's components, their biological activity, and its use in modern phytotherapy.
The effect of anise oil, dwarf-pine oil and chamomile oil against different thymidine-kinase-positive (aciclovir-sensitive) and thymidine-kinase-negative (aciclovir-resistant) herpes simplex virus type 1 (HSV-1) strains was examined. Clinical HSV-1 isolates containing frameshift mutations in the thymidine kinase (TK) gene, an insertion or a deletion, yield a non-functional thymidine kinase enzyme resulting in phenotypical resistance against aciclovir. The inhibitory activity of three different essential oils against herpes simplex virus isolates was tested in-vitro using a plaque reduction assay. All essential oils exhibited high levels of antiviral activity against aciclovir-sensitive HSV strain KOS and aciclovir-resistant clinical HSV isolates as well as aciclovir-resistant strain Angelotti. At maximum noncytotoxic concentrations of the plant oils, plaque formation was significantly reduced by 96.6-99.9%, when herpesviruses were preincubated with drugs before attachment to host cells. No significant effect on viral infectivity could be achieved by adding these compounds during the replication phase. These results indicate that anise oil, dwarf-pine oil and chamomile oil affected the virus by interrupting adsorption of herpesviruses and in a different manner than aciclovir, which is effective after attachment inside the infected cells. Thus the investigated essential oils are capable of exerting a direct effect on HSV and might be useful in the treatment of drug-resistant viruses. Chamomile oil did not reveal any irritating potential on hen's egg chorioallantoic membrane, demonstrated the highest selectivity index among the oils tested and was highly active against clinically relevant aciclovir-resistant HSV-1 strains.
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