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ORIGINAL CONTRIBUTION AND CLINICAL INVESTIGATIONMethods and MaterialsBacteria testedThe bacteria under study(Klebsiella pneumoniae, Proteusspp., Pseudomonas aeruginosa,Staphylococcus aureus andStreptococcus mutans) wereobtained from human infections fromHawlery ferkary hospital in Erbil city,Iraq. The isolates were inoculatedon nutrient agar to obtain singlecolonies. These were sub culturedon the same medium to check forthe purity of the isolated bacteria.Purified isolates were identifiedusing morphological, cultural andsome biochemical tests, as a moreaccurate method for identification.Essential OilsAll essential oils were obtained fromthe local market in Erbil city, Iraq, aslisted in Table (1).Plant ExtractionCollection and preparation of plantsamplesBoth plants (spearmint and seeds offenugreek) were obtained from themarket in Erbil city. For spearmint,the leaves were washed with tapwater, then with distilled water, thenleft for air drying until becomingcompletely dry, then both plantswere converted into powder formand stored in polyethylene bags inthe refrigerator at 4ºC for furtherprocessing.Extract preparationThe ethanolic extract of spearmintand seeds of fenugreek plantswas prepared by macerationmethod according to [23] with slightmodification, where 10 gm of plantpowder was steeped in 100mlethanol for three days then filtratedthrough eight layered muslin cloththen with filter paper (WhatmanNo.1), and centrifuged at 3000g for10 minutes; then the supernatantwas collected, and stored in a sterilebottle at 4ºC.Well diffusion techniqueScreening of antibacterial activity wasperformed by well diffusion technique[24]. The Nutrient agar (NA) plateswere seeded with 0.1 ml of theinoculums of each tested organism.The inoculums were spread evenlyover plate with a loop. A standardcork borer of 8 mm diameter wasused to cut uniform wells on thesurface of the NA and 100 µl ofessential oils or each concentrationof plant extracts was introduced inthe well. The plates were incubatedfor 24 hours at 37 ºC, and the zonesof inhibition were measured to thenearest millimeter (mm).Preparation of inoculumsTwo to three colonies from puregrowth of each tested organism weretransferred to 5 ml of nutrient broth.Broths were incubated overnight at37 ºC [25].Table 1: Plant oils14 MIDDLE EAST JOURNAL OF FAMILY MEDICINE VOLUME 10 ISSUE 7MIDDLE EAST JOURNAL OF FAMILY MEDICINE • VOLUME 7, ISSUE 10
ORIGINAL CONTRIBUTION AND CLINICAL INVESTIGATIONResults and DiscussionHistorically, many plants oils andextracts such as tea tree, myrrh andclove, have been used as topicalantiseptics, or have been reportedto have antimicrobial properties[26]. It is important to investigatescientifically those plants which havebeen used in traditional medicinesas potential sources of novelantimicrobial compounds [26].The antibacterial activity of oilof plants that were used in thisstudy is shown in Table (2). Theoil of pomegranate inhibited theStaphylococcus aureus, Proteus spp.and Pseudomonas aeruginosa,spearmint, pumpkin and rosemaryinhibited Staphylococcus aureus andProteus spp. and Staphylococcusaureus and Pseudomonasaeruginosa respectively, whilemustard, almond and radish inhibitedStaphylococcus aureus, but theother oils didn’t inhibit any of thebacteria under study. There are manyresearch studies that mention thatpomegranate is a strong antibacterialagent, and this may return to thepresence of phytocompounds in thephenols, tannins and flavonoids asmajor active constituents (95 percentpunicic acid; other constituents,including ellagic acid and sterols)may be responsible for theseactivities [12].Some researchers, such as [27, 28,10, 18, 19 and 29] mentioned thatparsley, seed oil of sesame, cuminand pumpkin have antibacterialactivity against G- and G+bacteria. The action mechanismof the essential oils is related totheir chemical composition andtheir antimicrobial activity is notattributable to a unique mechanismbut to a cascade of reactions in theentire bacterial cell. In fact, the actionmechanism of the essential oilsincludes: degradation of the cell wall,cytoplasmic-membrane damagingand cytoplasm coagulation, damageto membrane proteins, leakage ofcell contents, depletion of the* Values calculated as mean of triplicates.- No inhibition zone or less than 8 mm.Table 2: Antibacterial activity of essential oils of plantsMIDDLE MIDDLE EAST EAST JOURNAL JOURNAL OF OF FAMILY MEDICINE VOLUME • VOLUME 10 ISSUE 7, ISSUE 7 10 15
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Page 31 and 32: EDUCATION AND TRAININGlearning capa
Page 33 and 34: EDUCATION AND TRAININGpragmatism wh
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