Christine S. Evans

Antimicrobial action of essential oils : the effect of dimethylsulphoxide on the activity of cinnamon oil

Fifty‐one essential oils extracted from plants of known origin were tested for their antimicrobial activity against three bacteria, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and four yeasts, Torulopsis utilis, Schizosaccharomyces pombe, Candida albicans and Saccharomyces cerevisiae using the drop diffusion method. All showed antimicrobial activity against at least one of the micro‐organisms. Following this preliminary screening, 13 essential oils showing antimicrobial activity against at least five of the micro‐organisms were tested in the range 50 μg ml−1 to 500 μg ml−1 using broth micro dilution techniques with dimethylsulphoxide (DMSO) as a dispersing solvent. The concentration of most of the oils required for total inhibition of growth was >500 μg ml−1. Further studies on the antimicrobial action of cinnamon oil in the range 10–150 μg ml−1 showed that 50‐fold higher activity was found when no dispersing solvent was used.

Oxalate production by Basidiomycetes, including the white-rot species Coriolus versicolor and Phanerochaete chrysosporium

Oxalate was found to accumulate in liquid culture media from the growth of the white-rot basidiomycetes Coriolus versicolor, Heterobasidion annosum, Pleurotus florida and Phanerochaete chrysosporium. Whereas little oxalate accumulated during active growth, millimolar concentrations of oxalate were detected in culture media during the stationary phase. The basidiomycete Agaricus bisporus, the cultivated mushroom, also accumulated oxalate in its culture medium in the stationary phase. In comparison, the brown-rot fungi Amyloporia xantha, Coniophora marmorata, C. puteana and Poria vaporaria accumulated oxalate in the primary metabolic phase and throughout growth up to 35 days. Oxalate accumulation (0.04–10.0 mm) in white-rot cultures did not lower the pH of the medium during growth, whereas in brown-rot cultures oxalate (2.0–20.0 mm) reduced the media pH during growth. Cultures of Agaricus bisporus, C. puteana and Coriolus versicolor grown on solid media containing high levels of calcium (50 or 100 mm calcium chloride) produced calcium oxalate crystals to varying extents on the surface of the hyphae.

Cytotoxicity of clove (Syzygium aromaticum) oil and its major components to human skin cells.

Abstract.  The essential oil extracted from clove (Syzygium aromaticum) is used as a topical application to relieve pain and promote healing in herbal medicine and also finds use in the fragrance and flavouring industries. Clove oil has two major components, eugenol and β‐caryophyllene, which constitute 78% and 13% of the oil, respectively. Clove oil and these components are generally recognized as ‘safe’, but the in‐vitro study here demonstrates cytotoxic properties of both the oil and eugenol, towards human fibroblasts and endothelial cells. Clove oil was found to be highly cytotoxic at concentrations as low as 0.03% (v/v) with up to 73% of this effect attributable to eugenol. β‐caryophyllene did not exhibit any cytotoxic activity, indicating that other cytotoxic components may also exist within the parent oil.

Antimicrobial action of palmarosa oil (Cymbopogon martinii) on Saccharomyces cerevisiae

The essential oil extracted from palmarosa (Cymbopogon martinii) has proven anti-microbial properties against cells of Saccharomyces cerevisiae. Low concentrations of the oil (0.1%) inhibited the growth of S. cerevisiae cells completely. The composition of the sample of palmarosa oil was determined as 65% geraniol and 20% geranyl acetate as confirmed by GC-FTIR. The effect of palmarosa oil in causing K(+) leakage from yeast cells was attributed mainly to geraniol. Some leakage of magnesium ions was also observed. Blocking potassium membrane channels with caesium ions before addition of palmarosa oil did not change the extent of K(+) ion leakage, which was equal to the total sequestered K(+) in the cells. Palmarosa oil led to changes in the composition of the yeast cell membrane, with more saturated and less unsaturated fatty acids in the membrane after exposure of S. cerevisiae cells to the oil. Some of the palmarosa oil was lost by volatilization during incubation of the oil with the yeast cells. The actual concentration of the oil components affecting the yeast cells could not therefore be accurately determined.

Kava lactones and the kava-kava controversy.

Kava-kava is a traditional beverage of the South Pacific islanders and has had centuries of use without major side effects. Standardised extracts of kava-kava produced in Europe have led to many serious health problems and even to death. The extraction process (aqueous vs. acetone in the two types of preparations) is responsible for the difference in toxicity as extraction of glutathione in addition to the kava lactones is important to provide protection against hepatotoxicity. The Michael reaction between glutathione and kava lactones, resulting in opening of the lactone ring, reduces the side effects of the kava kava extracts. This protective activity was demonstrated using Acanthamoebae castellanii in which 100% cell death occurred with 100 mg ml(-1) kava lactones alone, and 40% cell death with a mixture of 100 mg ml (-1)glutathione and 100 mg ml (-1) kava lactones. A comparison of kava lactone toxicity with other pharmaceutical products is discussed and recommendations made for safe usage of kava-kava products

Reactions of pentachlorophenol with laccase from Coriolus versicolor.

Abstract Laccase, purified from Coriolus versicolor, removed pentachlorophenol (PCP) from solution at pH 5, depending on initial PCP concentration and amount of laccase. With 100 units of laccase, 100% of 25 μg ml−1 PCP and 60% of 200 μg ml−1 PCP were removed respectively over 72 h. No free chloride was released in the reaction. In reaction with 100 μg PCP, products were primarily polymers (about 80,000 MW) with only 2–3 pg of o- and p-chloranils formed. Polymers were stable to acid hydrolysis and no release of PCP, or other low-molecular-weight products, was detected over several weeks. Laccase has a potential use in the biotreatment of aqueous effluents containing PCP, with polymerised products being removed from solution due to their high molecular weight.

CYTOTOXICITY OF LAVENDER OIL AND ITS MAJOR COMPONENTS TO HUMAN SKIN CELLS

Abstract.  Lavender (Lavandula angustifolia) oil, chiefly composed of linalyl acetate (51%) and linalool (35%), is considered to be one of the mildest of known plant essential oils and has a history in wound healing. Concerns are building about the potential for irritant or allergenic skin reactions with the use of lavender oil. This study has demonstrated that lavender oil is cytotoxic to human skin cells in vitro (endothelial cells and fibroblasts) at a concentration of 0.25% (v/v) in all cell types tested (HMEC‐1, HNDF and 153BR). The major components of the oil, linalyl acetate and linalool, were also assayed under similar conditions for their cytotoxicity. The activity of linalool reflected that of the whole oil, indicating that linalool may be the active component of lavender oil. Linalyl acetate cytotoxicity was higher than that of the oil itself, suggesting suppression of its activity by an unknown factor in the oil. Membrane damage is proposed as the possible mechanism of action.

Effect of dissolved oxygen tension and ph on the production of extracellular protease from a new isolate of bacillus subtilis K2, for use in leather processing

Scale-up of production of an alkaline protease, previously characterised from a new isolate of Bacillus subtilis for use as a bating enzyme in leather processing, is described. Before large-scale commercial production of the protease is possible, characteristics of the growth of the bacterium and enzyme production in fermenters must be defined. In 2 dm3 fermenters an optimal specific activity of 296×103 U g −1 cell dry weight was obtained after 60 h with the dissolved oxygen tension (DOT) kept above 10% and pH left uncontrolled. Culture pH was 6 on inoculation, falling to 5.3 after 12 h before rising steadily to ∽8 at the end of fermentation. DOT was maintained above 10% by agitation in the range 300 to 500 rpm. The same criteria were adopted for scale-up to 20 dm3 but the increase in activity occurred 24 h later. © 1999 Society of Chemical Industry

Hydrogen-peroxide-producing system ofPleurotus eryngii involving the extracellular enzyme aryl-alcohol oxidase

The effect of benzyl alcohol, benzaldehyde and benzoic acid on the production of extracellular hydrogen peroxide (H2O2) by the ligninolytic fungusPleurotus eryngii was investigated. It was found that an equilibrium between oxidative and reductive reactions of these compounds is established, leading to the continuous production of H2O2. A multienzymatic cyclic system is proposed in which H2O2 is produced extracellularly by the action of aryl-alcohol oxidase on benzyl alcohol, the most abundant compound after redox reactions, and to a lower extent on benzaldehyde. The oxidation products of these reactions, benzaldehyde and benzoic acid, are reduced by intracellular dehydrogenases.

Localisation of degradative enzymes in white-rot decay of lignocellulose

The use of immunogold-cytochemical labelling techniques in electron microscopy of wood infected by basidiomycete fungi has assisted in the elucidation of the localisation of enzymes which degrade lignocellulose. The use of specific immunocytochemical techniques is discussed with respect to the authenticity and accuracy of the methods, the use of adequate controls in the gold-labelling procedure, and the immunospecificity of the antibodies.Localisation of the lignin-degrading enzymes, lignin-peroxidase and laccase, has shown that these enzymes do not bind to wood cell walls unless the process of decay has already commenced. Similarly localisation of cellulases Endoglucanase II (EGII) and Cellobiohydrolase I (CBHI) has shown that these enzymes only bind to exposed ends of cellulose fibrils and to partially degraded areas of the wood cell wall. β-Glucosidase is always immobilised within the extracellular polysaccharide layer surrounding fungal hyphae.This review postulates that there is regulation of the release sequence of these lignocellulolytic enzymes defining the spatial arrangement between the hyphae and the wood cell wall. This hypothesis is presented diagrammatically.