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The effect of essential oils on methicillin-resistant ...

The effect of essential oils on methicillin-resistantStaphylococcus aureususing a dressing modelV. Edwards-Jonesa,*, R. Bucka, Shawcrossa, Dawsona, K. DunnbaDepartment of Biological Sciences, the Manchester Metropolitan University, Chester Street, Manchester, M15GD, Centre, Acute Block, Wythenshawe Hospital, Southmoor Road, Manchester, M239LT, 4 June 2004 AbstractPatchouli, tea tree, geranium, lavender essential oils and CitricidalTM(grapefruit seed extract) were used singly and in combination toassess their anti-bacterial activity against three strains ofStaphylococcus aureus: OxfordS. aureusNCTC 6571 (Oxford strain), Epidemicmethicillin-resistantS. aureus (EMRSA 15)and MRSA (untypable). The individual essential oils , extracts and combinations wereimpregnated into filter paper discs and placed on the surface of agar plates, pre-seeded with the appropriate strain effects of the vapours of the oils and oil combinations were also assessed using impregnated filter paper discs that were placed on theunderside of the Petri dish lid at a distance of 8 mm from the bacteria.

essential oils and extracts were tested for their anti-bacterial properties using an invitro dressing model to assess the anti- ... All experiments were carried out three times. 2.4. In vitro dressing model Aliquots (100 ml) of essential oil and combinations that

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1 The effect of essential oils on methicillin-resistantStaphylococcus aureususing a dressing modelV. Edwards-Jonesa,*, R. Bucka, Shawcrossa, Dawsona, K. DunnbaDepartment of Biological Sciences, the Manchester Metropolitan University, Chester Street, Manchester, M15GD, Centre, Acute Block, Wythenshawe Hospital, Southmoor Road, Manchester, M239LT, 4 June 2004 AbstractPatchouli, tea tree, geranium, lavender essential oils and CitricidalTM(grapefruit seed extract) were used singly and in combination toassess their anti-bacterial activity against three strains ofStaphylococcus aureus: OxfordS. aureusNCTC 6571 (Oxford strain), Epidemicmethicillin-resistantS. aureus (EMRSA 15)and MRSA (untypable). The individual essential oils , extracts and combinations wereimpregnated into filter paper discs and placed on the surface of agar plates, pre-seeded with the appropriate strain effects of the vapours of the oils and oil combinations were also assessed using impregnated filter paper discs that were placed on theunderside of the Petri dish lid at a distance of 8 mm from the bacteria.

2 The most inhibitory combinations of oils for each strain were used in adressing model constructed using a four layers of dressings: the primary layer consisted of either JelonetTMor TelfaClearTMwith or withoutFlamazineTM; the second was a layer of gauze, the third a layer of Gamgee and the final layer was Crepe bandage. The oil combinations wereplaced in either the gauze or the Gamgee layer. This four-layered dressing was placed over the seeded agar plate, incubated for 24 h at 378 Cand the zones of inhibition measured. All experiments were repeated on three separate occasions. No anti-bacterial effects were observedwhen FlamazineTMwas smeared on the gauze in the dressing model. When TelfaclearTMwas used as the primary layer in the dressing modelcompared to JelonetTM, greater zones of inhibition were observed. A combination of CitricidalTMand geranium oil showed the greatest-anti-bacterial effects against MRSA, whilst a combination of geranium and tea tree oil was most active against the methicillin-sensitiveS.

3 Aureus(Oxford strain). This study demonstrates the potential of essential oils and essential oil vapours as antibacterial agents and for use in thetreatment of MRSA infection.#2004 Elsevier Ltd and ISBI. All rights :MRSA; Wound infection; Dressings; essential oils1. IntroductionDespite major advances in wound management, infectionstill remains an important factor in wound healing. In burns,approximately 75% of deaths are due to complications withsepsis resulting from wound infection[1]. Among otheradverse effects, infection delays healing, contributes to graftfailure and can increase the depth of a burn. Approximately30% of burn wounds become colonised with Staphylococcusaureus[2]and outbreaks of methicillin-resistantS. aureus(MRSA) have created major problems for burn units andintensive care units in terms of cross-infection and rehabi-litation of the patient due to imposed barrier nursing[3].

4 Some MRSA strains, termed epidemic MRSA (EMRSA),have the ability to spread rapidly among patients and thedominant clonal EMRSA types 15 and 16 are problematic inthe UK[4,5]. A new strain,EMRSA 17, has recently beendescribed in the UK[6]. Whilst vancomycin is one of the fewremaining effective systemic antibiotics available for treat-ment, resistance has been reported and there is major con-cern that total antibiotic-resistant strains may emerge in theimmediate future[7]. At present, topical anti-microbialtherapy is the single most important component of woundcare to prevent infection[8]and in hospitalised burnpatients, FlamazineTMis by far the most frequently 30 (2004) 772 777* Corresponding author. Tel.: +0161 247 1229; fax: +0161 247 (V. Edwards-Jones).0305-4179/$ #2004 Elsevier Ltd and ISBI. All rights prophylactic agent[9]but does not always penetrateinto the wound[10]and cannot be used to eradicate carriagefrom the patient or the therapies are being sought for treatment ofMRSA and one area of interest is the use of essential is susceptible to tea tree oil[11]but there areconcerns about its toxicity[12].

5 Tea tree oil has beenaccepted as a powerful antibacterial agent[13]and is soldcommercially in a wide range of products. There arehundreds of other essential oils available for use, many withknown antibacterial properties. These oils contain numerousconstituents that contribute to the characteristic odour andmedicinal effects. The major chemical components thataccount for the pleasant aromatic odours are primarilyterpenenes, monoterpenes and linalool[14]. The presenceand quantity of the various components varies between oilsand determines the individuality of the oil[15]. Althoughessential oils are known for their antimicrobial properties,medical teams rarely use them. This is primarily due to lackof scientific evidence of their efficacy, toxicity issues and theavailability of conventional therapy. In this study, a range ofessential oils and extracts were tested for their anti-bacterialproperties using an in vitro dressing model to assess the anti-microbial action of the Materials and Bacterial strainsThree strains ofS.

6 Aureuswere used: an antibiotic-susceptible strain,S. aureus NCTC 6571(Oxford strain); amethicillin-resistantS. aureus(non-typable) and a methi-cillin-resistantS. aureus(phage type15). The two strains ofMRSA were isolated from the wounds of two burn patientson the Burns Unit, Withington Hospital, Manchester, essential oilsFour essential oils were used in the study: tea tree(Melaleuca alternifolia), patchouli (Pogostemon cablin),lavender (Lavendula officinalis), geranium (Pelargoniumgraveolens) (Supplier: Essentially oils , Chipping Norton,UK); CitricidalTM(grapefruit seed extract), a commerciallyavailable antibacterial agent, was also Assessment of antibacterial action usingimpregnated filter paper discsAliquots (20ml) of each of the four oils or extract andtheir combinations (50:50) were spotted onto 5 mmfilterpaper discs and allowed to air dry for 40 min.

7 The discs werethen placed onto the surface of sensitivity test agar (STA)(Lab M, Bury, UK) that had been seeded with a suspensionof 105organisms/ml of each strain ofStaphylococcus(directcontact). Another set of seeded plates was exposed to thevapours of the oils or combinations by placing animpregnated disc in the centre of the lid of the Petri dish(vapour contact). All plates were incubated at 378C for 24 hand zones of inhibition (diameter in mm) measured on theagar surface. All experiments were carried out three In vitro dressing modelAliquots (100ml) of essential oil and combinations thatshowed antimicrobial activity in the disk diffusion experi-ments were placed onto a small central area of either theGamgee or gauze dressing layers. STA plates were seededwith a suspension of 105organisms/ml of each strain covered with four layers of dressings as shown inFig.

8 1. Modifications were made to the primary layer byconstructing a model with or without FlamazineTMandusing either Telfa ClearTMor JelonetTM, or both, which arereduced adherence dressings. The in vitro models were thenincubated at 378C for 24 h and the zones of inhibitionmeasured. All experiments were carried out three Antibacterial action of single oils : direct contactAll bacterial strains showed some susceptibility to eachindividual essential oil when tested in direct contact usingV. Edwards-Jones et al. / Burns 30 (2004) 772 777773 Fig. 1. The construction of the in vitro dressing disc diffusion method. The size of the zone of inhibitionvaried depending upon the oils and the strain ofS. aureusused. Tea tree oil gave the largest zones of inhibition whentested against MRSA than any of the other oils . MRSA appeared to be more susceptible to tea tree oil than theOxfordStaphylococcus.

9 Patchouli, lavender, geranium andCitricidalTMinhibited growth of the three strains ofstaphylococcus, but the OxfordStaphylococcusshowedlarger zones of inhibition (Fig. 2). Antibacterial action of single oils : vapour phaseThe OxfordStaphylococcuswas most susceptible to thevapours of tea tree, patchouli and geranium oil, but not tolavender and CitricidalTM. The MRSA strains showed nosusceptibility to the vapours apart from in one strain, wheretea tree oil showed limited effect (Fig. 3). Antibacterial action of oil combinations: directcontactPair-wise combinations of these oils were tested for theirantibacterial action. Where the disks were placed in directcontact with strains of staphylococcus,five of these com-binations had little or no effect on the antibacterial activity(patchouli and lavender, patchouli and CitricidalTM, patchouliand geranium, lavender and CitricidalTM, and geranium andCitricidalTM) compared to the single oil.

10 However, the otherfive combinations (patchouli and tea tree, lavender andgeranium, lavender and tea tree, grapefruit and tea tree andgeranium and tea tree) produced increased zones of inhibitionwith the OxfordStaphylococcus, but not in the MRSA the MRSA strains, two combinations (lavender and tea tree,CitricidalTMand tea tree) showed decreased activity whencompared to tea tree oil alone (Fig. 4). Antibacterial action of oil combinations:vapour phaseIn the vapour phase, there were marked differences inobserved antimicrobial effect of certain oil combinations;this was dependent upon the oil combination used and thestrain of staphylococcus. Increased zones of inhibition wereobserved with the two MRSA strains when combinations ofpatchouli and tea tree, CitricidalTMand geranium, andCitricidalTMand tea tree were used (Fig. 5). Whereas withsingle oils , no zones of inhibition or very small zones werenoted (Fig.)


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