FYP Proposal Draft

Development and Screening of Cinnamon Oil Based Topical Formulations for their Antibacterial Activity

Tee Lui Shi

Bachelor of PharmacySchool of PharmacyTaylors University

Supervised byDr. Manimaran Sellappan

Date

1.0 IntroductionEssential oils are aromatic, volatile mixtures of plant secondary metabolites, which cover a broad spectrum of activities thanks to their chemical diversity- its common to find 2060 constituents in an individual essential oil (Kalemba & Kunicka, 2003). Cinnamon oil is one of those essential oils that have received much attention. For thousands of years, cinnamon oil has been used as a spice due to its special aroma and now it has moved beyond the realm of taste as more and more studies demonstrate the medicinal properties of cinnamon oil (Bandara, Uluwaduge, & Jansz, 2012; Ranasinghe et al., 2013). Although there are about 300 species of Cinnamomum, only four species are used to obtain the spice cinnamon with Ceylon True cinnamon (Cinnamomum zeylanicum) and Chinese Cassia cinnamon (Cinnamomum aromaticum) as the most widely available varieties (Jayaprakasha & Rao, 2011). The purpose of present study is to developan effective topical antibacterial formulation of C. zeylanicum oil and to evaluate its antibacterial activity. The cream formulated will be the first cinnamon oil antibacterial cream targeted for human use. The rationale of choosing C. zeylanicum instead of C. aromaticum is because the antimicrobial activities of C. zeylanicum oil are well established and documented in the World Health Organization monograph (Who, 1999) compare to the corresponding scanty information for C. cassia (Ooi et al., 2006).

2.0 Literature review2.1 Phytochemical reviewThe plant organs of C. zeylanicum produce essential oil of varying chemical composition with the major constituents of its leaf being eugenol (76.74-90.2%) (Jantan, Karim Moharam, Santhanam, & Jamal, 2008; Paranagama et al., 2001; Singh, Maurya, deLampasona, & Catalan, 2007). For its bark, cinnamaldehyde (49.9-62.8%) is being reported as the primary constituent (Paranagama et al., 2001; Jantan, Karim Moharam, Santhanam, & Jamal, 2008; Simi et al., 2004; Singh, Maurya, deLampasona, & Catalan, 2007; Unlu, Ergene, Unlu, Zeytinoglu, & Vural, 2010)Camphor (47.42%) is being reported as the major constituent of C. zeylanicum root (Paranagama et al., 2001). Cinnamaldehyde and eugenol have also been reported as the major components of Cinnamomum spp. essential oil extract (Azeredo, Santos, Helena, Noronha, & Maia, 2014; Cheng, Liu, Hsui, & Chang, 2006; Usta, Kreydiyyeh, Barnabe, Bou-Moughlabay, & Nakkashchmaisse, 2003).This chemical diversity of C. zeylanicum has been accounted for its wide variety of medicinal benefits and usage in different industries (Ranasinghe et al., 2013). 2.2 Biological reviewBesides its traditional use as a spice in flavouring industry and as a rub to increase blood circulation in aromatherapy (Rana, Singh, & Gwal, 2011), modern studies from different parts of the world have demonstrate numerous beneficial health effects of C. zeylanicum, such as anti-inflammatory (Vetal, Bodhankar, Mohan, & Thakurdesai, 2013), anti-oxidant (Jayaprakasha & Rao, 2011; Singh et al., 2007) and antimicrobial (Naveed et al., 2013; Ooi et al., 2006)Anti-bacterial action of cinnamon oil is considered to arise mainly from the potential ofhydrophobic essential oil to disrupt the bacterial cell membrane and its structures which leads to ion leakage (Bakkali, Averbeck, Averbeck, & Idaomar, 2008; Burt, 2004; Rana et al., 2011). Many studies have shown that cinnamaldehyde is the primary compound responsible for major antibacterial activity of cinnamon oil (Rana et al., 2011; Unlu et al., 2010)Cinnamon oil alone or in combination with triclosan, gentamicin, or chlorhexidine can effectively inhibit biofilm formation, detach existing biofilms, and kill bacteria in biofilms of clinical Staphylococcus epidermidis strains without causing resistance (Hamidpour, Hamidpour, Hamidpour, & Shahlari, 2015).Cinnamon oil together with its main constituents- cinnamaldehyde (0.05%v/v) and eugenol (0.01%v/v) is reported to be effective against Pseudomonas aeruginosa and Escherichia coli as they can markedly inhibit the biofilm formation of these bacteria by 96% and 90% respectively (Kim, Lee, Kim, Baek, & Lee, 2015).Study done using disc diffusion method showed that essential oil of cinnamon fully inhibit the growth of four gram-positive and gram-negative bacterial strains, namely Pasturella multocida, Escherichia coli, Bacillus subtilis and Staphylococcus aureus (Saleem, Bhatti, Jilani, & Hanif, 2015).In a study done on cinnamon, geranium and lavender essential oils, cinnamon bark oil was the most active against clinical and environmental strains of Acinetobacter baumannii with MIC values ranging from 0.5 to 2.5 L/mL (Sienkiewicz et al., 2014).In a study done on essential oils of cinnamon, rosemary, thyme, clove and mint, cinnamon oil showed the strongest bactericidal effect against Escherichia coli (Kalaba & Kalaba, 2014).Essential oil obtained from C. zeylanicum inhibited activity of Staphylococcus, Enterococcus, Enterobacter and Acinetobacter genera even in low concentration, with the MIC for Gram-positive bacteria between 01.25-1.5 L/mL and for Gram-negative between 1.0-1.75 L/mL. No sign of resistance as those seen with the recommended conventional antibiotics was observed (Urbaniak, Gowacka, Kowalczyk, Lysakowska, & Sienkiewicz, 2014).Among all four selected essential oils, oil from the bark of C. zeylanicum showed best antibacterial activity against all tested multidrug-resistance strains in the MIC assay, namely, Salmonella typhi (D1 Vi-positive), Salmonella typhi (G7 Vi-negative), Salmonella paratyphi, Escherichia coli, Staphylococcus aureus, Pseudomonas fluorescens and Bacillus licheniform with the most prominent ones being 2.9 mg/mL concentration against S typhi G7 Vi-negative and P fluorescens strains (Naveed et al., 2013).C. zeylanicum essential oil suppressed the growth of Acinetobacter spp. and exerted synergistic effect with the antibiotic amikacin by drastically reducing the MIC of amikacin when its sub-inhibitory concentrations at 78.125g/mL (MIC/8) is added to the growth medium (Guerra et al., 2012). In a study involving passaging bacteria up to fifty times with sequential exposure of cinnamon essential oils, no resistance was reported out of the 48 clinical isolates and 12 reference strains under study (Becerril, Nern, & Gmez-Lus, 2012).C. zeylanicum showed the highest activity among 19 essential oils when screened against four bacteria strains namely Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhimurium and Bacillus subtilis, with Staphylococcus aureus and B. subtilis being most susceptible, exhibiting MIC values 0.2 and 0.4 L oil/well respectively (Rana et al., 2011).Commercially available sugar-sweetened cinnamon chewing gum may benefit halitosis by reducing volatile sulfur compounds producing anaerobes in the oral cavity as there was significant reductions in total salivary anaerobes (p < 0.01) and H2S-producing salivary anaerobes (p < 0.01) after subjects chewed gum containing cinnamic aldehyde and natural flavors (Zhu, Carvalho, Scher, & Wu, 2011).Among 21 essential oils tested, cinnamon oil was the most effective antibacterial agent with MIC values ranging from 0.8 to 3.2 mg/mL to show maximum activity against P. aeruginosa, B. subtilis, P. vulgaris, K. pneumonia and S. aureus (Prabuseenivasan, Jayakumar, & Ignacimuthu, 2006).Potential use of Cinnamomum zeylanicum bark oil in the formula of natural remedies for the topical treatment of infections is demonstrated in a study by using disc diffusion (DD) and minimum inhibitory concentration (MIC) methods. The essential oil showed strong antibacterial activity against all 21 bacteria species tested and was highly effective against Gram positive Staphylococcus, Streptococcus, Enterococus and Gram negative Pseudomonas aeruginosa with a MIC of 40mm(Unlu et al., 2010).Cinnamomum zeylanicum essential oils inhibited growth of Salmonella typhi, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Bacillus subtilis with gram-negative organisms being more susceptible than gram-positive ones (Dubey, Rana, & Shukla, 2005).Essential oil obtained from the stem-bark of Cinnamomum species exhibit significant antibacterial activity against a wide range of Gram positive and Gram negative bacteria, especially Methicillin-resistant Staphylococcus aureus (MRSA) with zone of inhibition up to 21.0000 1.4142mm (Buru, Pichika, Neela, & Mohandas, 2014) *not c.zeylanicum, but got cinnamaldehyde*

3.0 Objective To develop a new antibacterial topical formulation for human use using C. zeylanicum essential oils. To provide an alternative for the treatment of infections caused by resistant and multi-resistant bacteria.

4.0 ScopeThe rising of antibiotic resistant bacteria in the recent decades has become a serious threat to public health worldwide (Handali, Hosseini, Ameri, & Moghimipour, 2011; Warnke et al., 2009). This resistance towards antibiotic is fuelled by its widespread misuse, leading to insufficiency of antibiotic therapies that are effective. There is a strong need for the development of new antibiotic, however, designing and developing new drugs takes a lot of time and money. This has caused many major pharmaceutical companies to abandon the field, especially when the recuperation rate of R&D expenses are low due to rapid emergence of bacterial resistance shortly after the drug is marketed. If this crisis situation persists, experts predict that a pre-antibiotic era will return, where patients may routinely die from seemingly minor infections (Qureshi, Agrawal, Madan, Pandey, & Chauhan, 2015). Thus, a different alternative should be explored, that being the essential oils which affect a wide range of antibiotic resistant microbes.

5.0 Research Methodology5.1 Collection and identification of essential oilsThe essential oil of C. zeylanicum is to be purchased from Malaysian supplier in May 2015. Identity and quality of the essential oil is verified through the specifications in certificates of analysis provided by the suppliers. 5.2 Development of formulationsThis product will be the first antibacterial cinnamon oil cream that is targeted for human use. Cream formulation is chosen as topical formulation is said to be advantageous in the aspects of solubility, concentration of drug released and delivery of drug directly to the site of action (Okore, Attama, Ofokansi, Esimone, & Onuigbo, 2011; Woodruff J, 1995). On the other hand, cream formulation is easily spreadable and washable with water which facilitate its application (Muller et al., 2003; Okore et al., 2011; Orafidiya, Oyedele, Shittu, & Elujoba, 2001; Sellappan M., 2014). The base cream contains water and oil phases, suitable excipients are added to the formulations besides the cinnamon oil. The tentative formula of the cream is illustrated in the tables below.Table 1: Tentative formula of the 2% cream formulation (100g)Ingredient% w/w

1.Stearic acid16.00

2.Potassium hydroxide02.00

3.Cetosteryl alcohol05.00

4.Liquid paraffin03.50

5.Petroleum jelly03.50

6.Methyl paraben sodium00.16

7.Propyl paraben sodium00.04

8.Glycerin07.00

9.Cinnamon oil02.00

10.Purified water60.80

Table 2: Tentative formula of the 5% cream formulation (100g)Ingredient% w/w

1.Stearic acid16.00







8.Glycerin07.00

9.Cinnamon oil05.00


Table 2: Tentative formula of the 10% cream formulation (100g)Ingredient% w/w

1.Stearic acid16.00







8.Glycerin07.00

9.Cinnamon oil10.00


5.3 ProcedureAqueous cream seems to be as effective as the test agents because of its easy penetration through the skin (Handali et al., 2011; Muller et al., 2003; Orafidiya et al., 2001). Stearic acid, cetosteryl alcohol, liquid paraffin and petroleum jelly to be melted on a steam bath at 75oC. The remaining ingredients are to be dissolved in water and boiled at 75oC. This aqueous solution to be added to the above oily phase with agitation. Potassium hydroxide reacts with a portion of stearic acid to form potassium stearate, which emulsifies the unreacted stearic acid as dispersed phase. Excessive agitation to be avoided so as to avoid air getting entrapped within the cream. 2% and 5% of the essential oils are to be mixed uniformly with the above prepared cream base individually and combination of both the oils with equal proportions using a mechanical stirrer. Glycerin to be added finally and mixed. The formulated creams are filled in a suitable plastic container.

5.4 Evaluation of formulationsThe prepared cream formulations are to be subjected to various physical evaluations listed below (Baboota et al., 2011; Okore et al., 2011). 5.4.1 Physical EvaluationA) Physical appearanceThe formulated creams are observed visually for their visual appearance, transparency, colour, consistency and feel upon application such as stickiness, smoothness and cooling effects.B) Determination of pH The pH of the prepared cream formulations and the base are to be determined using a pH meter (Eutech cyber scan pH 310) by preparing 1 gm of cream formulation to dissolve in 100 ml of water and maintaining a temperature of 25oC. The averages of the triplicate observations will be recorded.C) Determination of ExtrudabilityExtrudability is a useful empirical test to measure the force required to extrude the material from a tube. Since packaging in collapsible tubes has gained considerable importance in delivery of the desired quantity of cream, measurement of extrudability becomes an important critical data for creams. In the present study, the method adopted for evaluating cream formulation for extrudability will be based upon the quantity (in percentage) of the cream extruded from a collapsible tube on application of certain load. The more the quantity extruded, the better is the extrudability.

The cream formulation to be filled in a standard capped collapsible sealed tube. The tube will be weighed and recorded. The tube will be placed between two-glass slides and is clamped. A 500 g weight to be placed over the glass slide and then the cap is opened. The amount of cream extruded is collected and weighed. The percentage of cream extruded will be calculated and recorded as per grades allotted.D) Determination of Viscosity:The viscosity of formulated cream bases was determined. The viscosity determinations were carried out on Brookfield viscometer using spindle number S06 and the determinations were carried out in triplicate and the average of three reading is recorded. Antibacterial assayThe cylinder plate assay of drug potency is based on the measurement of the diameter of zones of microbial growth inhibition surrounding the cylinder (cups) containing various dilutions of test compounds. The following Gram-positive and Gram-negative bacteria will be used for the present study to determine the antimicrobial activity (Manimaran, Themozhil, Nanjan, & Suresh, 2007) (Baker and Breach, 1980; James et al., 1992).

5.5 ProcedurePetri dishes are to be cleaned, dried and sterilized. Then they are filled with a Nutrient Agar or SDA medium with uniform thickness. After solidifying, the plates are to be inoculated with the above mentioned organisms of Gram positive, Gram negative bacteria and fungi. In the first plate, five holes to be made in the inoculated plates by means of aluminum or stainless steel sterile borer with a height of 10 cm and an internal diameter of 6-8 mm. To the first, second and third hole, required quantity of 2% cream of oregano oil, geranium oil and mixed cream formulations to be introduced. The standard drug and cream base to be filled in the fourth and fifth hole. The same procedure will be followed for 5% cream formulations. Then the plates are to be kept in the refrigerator for 2 hrs for diffusion. All the plates are to be incubated at 37oC for 24 hrs. All the standards to be inoculated separately for bacteria and fungi.. The zone of inhibition was measured by using antibiotic zone reader.

6.0 OutcomeThis research project focuses on the three Ps: people, planet and profits. Thus, its outcome can be simplified as triple bottom. The outcomes are categorized into direct outcomes and indirect outcomes.6.1 Direct outcome A new antibacterial formulation consisting of cinnamon oil will be developed. There is currently no product for human use being marketed which uses cinnamon oil as the main active ingredient into a formulation. There will be more alternatives to treat an infection caused by MRSA strains.6.2 Indirect outcome It encourages continuous research and development in the field of essential oils. Potential anticancer, antidiabetic, anti-inflammatory and other drugs can be developed.

7.0 Timeframe DurationActivity2014

AprMayJuneJulyAugSeptOctNov

1Literature search

2Proposal writing

3Proposal presentation

4Collection of materials

5Formulation development

6Physical & biological evaluation of the formulation

7Data analysis

8Thesis writing

9Thesis presentation

8.0 BudgetNoDescriptionQuantity required (kg)Cost (RM)

1.Stearic acid0.100







8.Glycerin\400

9.Cinnamon oil50ml200

10.Nutrient agar- (to prepare media)0.7503400

11.Sabourad dextrose agar (to prepare media)

They can contain up to 2060 constituents at various concentrations, but usually only 23 main constituents that are responsible for their aromatic and biological properties (Kalemba & Kunicka, 2003).

9.0 ReferencesAzeredo, C. M. O., Santos, T. G., Helena, B., Noronha, L. De, & Maia, S. (2014). In vitro biological evaluation of eight different essential oils against Trypanosoma cruzi , with emphasis on Cinnamomum verum essential oil, 18.Baboota, S., Sharma, S., Kumar, A., Alam, M. S., Sahni, J., & Ali, J. (2011). Nanocarrier-based hydrogel of betamethasone dipropionate and salicylic acid for treatment of psoriasis. International Journal of Pharmaceutical Investigation, 1(3), 139. doi:10.4103/2230-973X.85963Bakkali, F., Averbeck, S., Averbeck, D., & Idaomar, M. (2008). Biological effects of essential oils - A review. Food and Chemical Toxicology, 46(2), 446475. doi:10.1016/j.fct.2007.09.106Bandara, T., Uluwaduge, I., & Jansz, E. R. (2012). Bioactivity of cinnamon with special emphasis on diabetes mellitus: A review. International Journal of Food Sciences and Nutrition, 63(3), 380386. doi:10.3109/09637486.2011.627849Becerril, R., Nern, C., & Gmez-Lus, R. (2012). Evaluation of bacterial resistance to essential oils and antibiotics after exposure to oregano and cinnamon essential oils. Foodborne Pathogens And Disease, 9(8), 699705. doi:10.1089/fpd.2011.1097Burt, S. (2004). Essential oils: Their antibacterial properties and potential applications in foods - A review. International Journal of Food Microbiology, 94(3), 223253. doi:10.1016/j.ijfoodmicro.2004.03.022Buru, A. S., Pichika, M. R., Neela, V., & Mohandas, K. (2014). In vitro antibacterial effects of Cinnamomum extracts on common bacteria found in wound infections with emphasis on methicillin-resistant Staphylococcus aureus. Journal of Ethnopharmacology, 153(3), 587595. doi:10.1016/j.jep.2014.02.044Cheng, S.-S., Liu, J.-Y., Hsui, Y.-R., & Chang, S.-T. (2006). Chemical polymorphism and antifungal activity of essential oils from leaves of different provenances of indigenous cinnamon (Cinnamomum osmophloeum). Bioresource Technology, 97, 306312. Retrieved from 10.1016/j.biortech.2005.02.030Dubey, R. C., Rana, A., & Shukla, R. K. (2005). Antibacterial Activity of Essential Oils of Some Medicinal Plants Against Certain Human Pathogens. INDIAN DRUGS -BOMBAY-, 42(7), 443446. Retrieved from http://ezproxy.taylors.edu.my/login?url=http://search.ebscohost.com/login.aspx?direct=true&db=edsbl&AN=RN172549316&site=eds-live&scope=siteGuerra, F. Q. S., Mendes, J. M., Sousa, J. P. de, Morais-Braga, M. F. B., Santos, B. H. C., Melo Coutinho, H. D., & Lima, E. de O. (2012). Increasing antibiotic activity against a multidrug-resistant Acinetobacter spp by essential oils of Citrus limon and Cinnamomum zeylanicum. Natural Product Research, 26(23), 22352238. doi:10.1080/14786419.2011.647019Hamidpour, R., Hamidpour, M., Hamidpour, S., & Shahlari, M. (2015). Cinnamon from the selection of traditional applications to its novel effects on the inhibition of angiogenesis in cancer cells and prevention of Alzheimers disease, and a series of functions such as antioxidant, anticholesterol, antidiabetes, antibacterial, antifungal, nematicidal, acaracidal, and repellent activities. Journal of Traditional and Complementary Medicine, 5, 6670. doi:10.1016/j.jtcme.2014.11.008Handali, S., Hosseini, H., Ameri, a, & Moghimipour, E. (2011). Formulation and evaluation of an antibacterial cream from Oxalis corniculata aqueous extract. Jundishapur Journal of Microbiology, 54(11), 48254832.Jantan, I. Bin, Karim Moharam, B. A., Santhanam, J., & Jamal, J. A. (2008). Correlation Between Chemical Composition and Antifungal Activity of the Essential Oils of Eight Cinnamomum Species. Pharmaceutical Biology, 46(6), 406412. doi:10.1080/13880200802055859Jayaprakasha, G. K., & Rao, L. J. M. (2011). Chemistry, biogenesis, and biological activities of Cinnamomum zeylanicum. Critical Reviews in Food Science and Nutrition, 51(6), 54762. doi:10.1080/10408391003699550Kalaba, V., & Kalaba, D. (2014). COMPARATIVE EFFECTS OF ESSENTIAL OILS ON GROWTH OF Escherichia coli, 59.Kalemba, D., & Kunicka, a. (2003). Antibacterial and antifungal properties of essential oils. Current Medicinal Chemistry, 10(10), 813829. doi:10.2174/0929867033457719Kim, Y.-G., Lee, J.-H., Kim, S.-I., Baek, K.-H., & Lee, J. (2015). Cinnamon bark oil and its components inhibit biofilm formation and toxin production. International Journal of Food Microbiology, 195, 3039. doi:10.1016/j.ijfoodmicro.2014.11.028Manimaran, S., Themozhil, S., Nanjan, M. J., & Suresh, B. (2007). Chemical Composition and Antimicrobial Activity of Cone Volatile oil of Cupressus macrocarpa Hartwig from Nilgiris, India. NATURAL PRODUCT SCIENCES, 13(4), 279282. Retrieved from http://ezproxy.taylors.edu.my/login?url=http://search.ebscohost.com/login.aspx?direct=true&db=edsbl&AN=RN222528482&site=eds-live&scope=siteMuller, M. J., Hollyoak, M. A., Moaveni, Z., Brown, T. L. H., Herndon, D. N., & Heggers, J. P. (2003). Retardation of wound healing by silver sulfadiazine is reversed by Aloe vera and nystatin. Burns, 29, 834836. Retrieved from 10.1016/S0305-4179(03)00198-0Naveed, R., Hussain, I., Tawab, A., Tariq, M., Rahman, M., Hameed, S., Iqbal, M. (2013). Antimicrobial activity of the bioactive components of essential oils from Pakistani spices against Salmonella and other multi-drug resistant bacteria. BMC Complementary and Alternative Medicine, 13(1), 265. doi:10.1186/1472-6882-13-265Okore, V. C., Attama, a a, Ofokansi, K. C., Esimone, C. O., & Onuigbo, E. B. (2011). Formulation and evaluation of diacerein cream. Indian Journal of Pharmaceutical Sciences, 73(3), 3238. doi:10.4103/0250-474X.93515Ooi, L. S. M., Li, Y., Kam, S.-L., Wang, H., Wong, E. Y. L., & Ooi, V. E. C. (2006). Antimicrobial activities of cinnamon oil and cinnamaldehyde from the Chinese medicinal herb Cinnamomum cassia Blume. The American Journal of Chinese Medicine, 34(3), 511522. doi:10.1142/S0192415X06004041Orafidiya, L. O., Oyedele, a. O., Shittu, a. O., & Elujoba, a. a. (2001). The formulation of an effective topical antibacterial product containing Ocimum gratissimum leaf essential oil. International Journal of Pharmaceutics, 224(1-2), 177183. doi:10.1016/S0378-5173(01)00764-5Paranagama, P. a., Wimalasena, S., Jayatilake, G. S., Jayawardena, a. L., Senanayake, U. M., & Mubarak, a. M. (2001). A comparison of essential oil constituents of bark, leaf, root and fruit of cinnamon (cinnamomum zeylanicum blum) grown in Sri Lanka. Journal of the National Science Foundation of Sri Lanka, 29(3-4), 147153. doi:10.4038/jnsfsr.v29i3-4.2613Prabuseenivasan, S., Jayakumar, M., & Ignacimuthu, S. (2006). In vitro antibacterial activity of some plant essential oils. BMC Complementary and Alternative Medicine, 6, 39. doi:10.1186/1472-6882-6-39Rana, I. S., Singh, A., & Gwal, R. (2011). In vitro study of antibacterial activity of aromatic and medicinal plants essential oils with special reference to cinnamon oil. International Journal of Pharmacy and Pharmaceutical Sciences, 3(4), 376380.Ranasinghe, P., Pigera, S., Premakumara, G. a S., Galappaththy, P., Constantine, G. R., & Katulanda, P. (2013). Medicinal properties of true cinnamon (Cinnamomum zeylanicum): a systematic review. BMC Complementary and Alternative Medicine, 13(1), 275. doi:10.1186/1472-6882-13-275Saleem, M., Bhatti, H. N., Jilani, M. I., & Hanif, M. A. (2015). Bioanalytical evaluation of Cinnamomum zeylanicum essential oil. Natural Product Research, (May 2015), 13. doi:10.1080/14786419.2014.1002088Sellappan M., N. and P. T. K. (2014). DEVELOPMENT AND SCREENING OF TOPICAL HERBAL CREAM, 5(5), 383388.Sienkiewicz, M., Gowacka, A., Kowalczyk, E., Wiktorowska-Owczarek, A., Jwiak-Bbenista, M., & ysakowska, M. (2014). The Biological Activities of Cinnamon, Geranium and Lavender Essential Oils. Molecules, 19(12), 2092920940. doi:10.3390/molecules191220929Simi, A., Sokovi, M. D., Risti, M., Gruji-Jovanovi, S., Vukojevi, J., & Marin, P. D. (2004). The chemical composition of some Lauraceae essential oils and their antifungal activities. Phytotherapy Research: PTR, 18(9), 713717. Retrieved from http://ezproxy.taylors.edu.my/login?url=http://search.ebscohost.com/login.aspx?direct=true&db=cmedm&AN=15478207&site=eds-live&scope=siteSingh, G., Maurya, S., deLampasona, M. P., & Catalan, C. a N. (2007). A comparison of chemical, antioxidant and antimicrobial studies of cinnamon leaf and bark volatile oils, oleoresins and their constituents. Food and Chemical Toxicology, 45(9), 16501661. doi:10.1016/j.fct.2007.02.031Unlu, M., Ergene, E., Unlu, G. V., Zeytinoglu, H. S., & Vural, N. (2010). Composition, antimicrobial activity and in vitro cytotoxicity of essential oil from Cinnamomum zeylanicum Blume (Lauraceae). Food and Chemical Toxicology, 48(11), 32743280. doi:10.1016/j.fct.2010.09.001Urbaniak, A., Gowacka, A., Kowalczyk, E., Lysakowska, M., & Sienkiewicz, M. (2014). [The antibacterial activity of cinnamon oil on the selected gram-positive and gram-negative bacteria]. Medycyna Dowiadczalna I Mikrobiologia, 66(2), 131141. Retrieved from http://search.ebscohost.com/login.aspx?direct=true&db=mdc&AN=25369660&site=ehost-liveUsta, J., Kreydiyyeh, S., Barnabe, P., Bou-Moughlabay, Y., & Nakkashchmaisse, H. (2003). Comparative study on the effect of cinnamon and clove extracts and their main components on different types of ATPases. Human & Experimental Toxicology, 22(7), 355362. Retrieved from 10.1191/0960327103ht379oaVetal, S., Bodhankar, S. L., Mohan, V., & Thakurdesai, P. a. (2013). Anti-inflammatory and anti-arthritic activity of type-A procyanidine polyphenols from bark of Cinnamomum zeylanicum in rats. Food Science and Human Wellness, 2(2), 5967. doi:10.1016/j.fshw.2013.03.003Warnke, P. H., Becker, S. T., Podschun, R., Sivananthan, S., Springer, I. N., Russo, P. a J., Sherry, E. (2009). The battle against multi-resistant strains: Renaissance of antimicrobial essential oils as a promising force to fight hospital-acquired infections. Journal of Cranio-Maxillofacial Surgery, 37(7), 392397. doi:10.1016/j.jcms.2009.03.017Who. (1999). WHO monographs on selected medicinal plants. World Health, 1(March), 390. Retrieved from http://kamillaviragzat.hu/wtDocument/browse/root/szakmai-anyagok/WHO_Monographs_vol1.pdfWoodruff J. (1995). Preservatives to fight the growth of mould. MANUFACTURING CHEMIST -LONDON-, 66(9), 34. Retrieved from http://ezproxy.taylors.edu.my/login?url=http://search.ebscohost.com/login.aspx?direct=true&db=edsbl&AN=EN030929445&site=eds-live&scope=siteZhu, M., Carvalho, R., Scher, A., & Wu, C. D. (2011). Short-term germ-killing effect of sugar-sweetened cinnamon chewing gum on salivary anaerobes associated with halitosis. The Journal Of Clinical Dentistry, 22(1), 2326. Retrieved from http://search.ebscohost.com/login.aspx?direct=true&db=mdc&AN=21290983&site=ehost-live

Cinnamomum zeylanicumBlume, Lauraceae, has many biological properties as analgesic, antiseptic, antispasmodic, aphrodisiac, astringent, carminative, haemostatic, insecticidal and parasiticide. Barks from branches, without the epidermis and subereous layer, is marketed as the commercial cinnamon which has long use in perfumery, culinary and native medicine fields (2,20). Previous research has revealed interesting antimicrobial effect inC. zeylanicumessential oil (10,27,28). Camphene, linalool,a-phelendrene,a-terpinene, limonene,b-cymene,a-cariophyllene, cinnamaldheyde and eugenol are some of the compounds found inC. zeylanicumessential oil (24,44).The present study aimed to evaluate the effect ofC. zeylanicumBlume essential oil on the growth and morphogenesis of someAspergillusspecies known as potential etiological agent of fungal infections.

http://www.dermnetnz.org/bacterial/ Skin bacteria Staphylococcus aureus Folliculitis Furunculosis(boils) andabscesses Impetigo(school sores) Methicillin (meticillin) resistant Staph. aureus Staphylococcal scalded skin syndrome Toxic shock syndrome Tropical pyomyositis Botryomycosis (pyoderma vegetans) Streptococcus pyogenes Cellulitis Erysipelas Impetigo Necrotising fasciitis Infectious gangrene Scarlet fever Rheumatic fever, erythema marginatum Overgrowth ofCorynebacteriumspp. (erythrasma,pitted keratolysis&trichomycosis axillaris)Less commonly, other bacteria may also cause infection with skin signs. These include: Neisseriaspecies, cause ofgonorrhoeaandmeningococcal disease Erysipelothrix insidiosa, cause oferysipeloid(usually an animal infection) Haemophilusspecies, cause ofchancroidandcellulitisin young children Helicobacter pylori, a stomach infection, which may be associated with some cases of chronicurticariaandrosacea Klebsiella rhinoscleromatis, cause ofrhinoscleroma Mycoplasma pneumoniae, a cause of pneumonia, causes non-specific erythema, bullous eruptions, urticarial rashes,erythema multiforme, mucositis and rarely,SJS/TEN Pseudomonas aeruginosacauses wound infections,athlete's foot,gram negative folliculitis, chronicparonychia(green nail syndrome),spa pool folliculitisandecthyma gangrenosum Calymmatobacterium granulomatis, cause ofgranuloma inguinale Bacillus anthracis, cause ofanthrax Clostridium perfringensand other species cause gas gangrene Treponemaspecies causesyphilis,yawsandpinta Bartonellaspecies causecat scratch fever,bacillary angiomatosisand bartonellosis Mycobacteriumspecies causetuberculosis,leprosyandatypical mycobacterial infectionsincludingBuruli ulcer Leptospira, cause ofleptospirosis, which may cause bleeding into the skin (purpura) Nocardia, cause of nocardiosis Yersinia pestis, cause of bubonicplague, which causes swollen lymph glands and pustules, ulcers and scabs on the skin Serratia marcescens, a facultative anaerobic gram-negative bacillus that may rarely cause skin infections such as cellulitis,abscessesand ulcers; usually in patients withimmunodeficiency. Fusibacteriumspecies,Bacillus fusiformis,Treponema vincentiand other bacteria may result intropical ulcer Burkholderiaspecies, cause ofmelioidosis and glanders, in which abscesses may be associated with systemic symptoms. Actinomcyesspecies, cause ofactinomycosis, in which granular bacteriosis occurs i.e. abscesses and sinus tracts draining sulphur-yellow granules. Vibrio vulnificus, a cause of septic shock characterised by blood-filled blisters. Brucellaspecies, cause ofbrucellosis, a febrile illness caught from unvaccinated animals or their unpasteurised milk. Salmonella species, particularlyS typhi(typhoid fever) Aeromonasfound in water rarely causes skin and soft tissue infections

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