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International Journal of Food Science and Technology 2009, 44, 11761182

Original article Chemical composition and inhibitory effect of essential oil and organic extracts of Cestrum nocturnum L. on food-borne pathogens
Sharif M. Al-Reza, Atiqur Rahman & Sun Chul Kang*
Department of Biotechnology, Daegu University, Kyoungsan, Kyoungbook 712-714, Korea (Received 14 October 2008; Accepted in revised form 28 Jan 2009)

Summary

In this study, we examined the chemical compositions of essential oil and tested the ecacy of oil and organic extracts of Cestrum nocturnum L. against food-borne pathogens. The chemical compositions of the oil was analysed by gas chromatography-mass spectrometry (GC-MS). Forty-seven compounds representing 93.28% of the total oil were identied. The oil [5 lL of 1:5 (v v) dilution of oil with methanol] and organic extracts of hexane, chloroform, ethyl acetate and methanol (300 lg per disc) of C. nocturnum displayed a great potential of antibacterial activity against Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes (ATCC 19166 and ATCC 15313), Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella typhimurium KCTC 2515 and Escherichia coli ATCC 8739. Also the oil had strong detrimental eect on the viable count of the tested bacteria. The results obtained from this study may contribute to the development of new antimicrobial agents with potential applications in food industries as natural preservatives to control food-borne pathogens.
Antibacterial activity, benzyl alcohol, Cestrum nocturnum L., essential oil, food-borne pathogens, phenylethyl alcohol.

Keywords

Introduction

Global interest in bio-preservation of food systems has recently been increased because of great economic costs of deterioration and poisoning of food products by food pathogens and is often responsible for the loss of quality and safety. Concern over pathogenic and spoilage micro-organisms in foods is increasing because of the increase in outbreaks of food borne disease. Currently there is a growing interest to use natural antibacterial compounds, like extracts of herbs and spices for the preservation of foods. Plant-derived essential oils and extracts of various species of edible and medicinal plants, herbs, and spices have long been used as natural agents for food preservation in food and beverages because of the presence of antimicrobial compounds (Nychas et al., 2003). In general, plant derived essential oils are considered as non-phytotoxic compounds and potentially eective against micro-organisms. In this context, the identication and evaluation of natural products for the control of these pathogens, to assure consumers a safe, wholesome, and nutritious food supply, can be considered an important international challenge.
* Correspondent: Fax: +82 53 850-6559; e-mail: sckang@daegu.ac.kr

With the increase of bacterial resistance to antibiotics, there is considerable interest to investigate the antimicrobial eects of essential oils and dierent extracts against a range of bacteria, to develop other classes of natural antimicrobials useful for the infection control or for the preservation of food (Bakri & Douglas, 2005). The Gram positive bacterium Staphylococcus aureus is mainly responsible for post-operative wound infections, toxic shock syndrome, endocarditis, osteomyelitis and food poisoning (Mylotte et al., 1987). Listeria monocytogenes is responsible for the severe food-borne illness, listeriosis, which has been recognised to be one of the emerging zoonotic diseases during the last two decades (Farber, 2000). The Gram negative bacterium Escherichia coli is present in human intestines and causes urinary tract infection, coleocystitis or septicemia (Singh et al., 2000). Cestrum nocturnum L. is a garden shrub from the family Solanaceae, the owers of which exude a special sweet fragrance at night, the main reason for its folk names night cestrum, lady of the night, night-blooming jessamine, and night blooming jasmine. It is widely naturalised in tropical and subtropical regions throughout the world, including Australia, Southern China and the Southernmost United States. It is also cultivated in Bangladesh in home yards and gardens. Several phytochemical studies have demonstrated the presence of important bioactive compounds in dierent parts of the

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plant: alkaloids, avonol glycosides, steroidal saponins, fatty acids, essential oils, phenols, and others (Bouchbaver et al., 1995). Practitioners use the plant externally for skin disorders, but several scientic reports demonstrate that it exhibits a wide spectrum of pharmacological activity when administered systemically or in isolated organ preparations. For example, it is used to treat arterial hypotension and as an analgesic, abortive, diuretic, antispasmodic, dyspeptic, antiviral, and smooth muscle relaxant; it also has negative inotropic and chronotropic actions (Perez-Saad & Buznego, 2008). Most of the species of Cestrum have found several applications in folk medicine. Cestrum parqui is used in Chilean folk medicine as antifebrile and for the treatment of fever and inammation. Chinese people use leaves of C. nocturnum for their pharmacological signicance in burns and swellings. It is also used for treating epilepsy and as stupefying charm medicine in West Indian Islands. The volatile oil of the species is known to be mosquito repellent and hence C. nocturnum and C. diurnum are used to prevent malaria in several African Nations (Ntonifor et al., 2006). The plants of the genus have further found use in perfumery, as ornamental plants, oral scent production etc. However, there is no report available in the literature on the analyses of essential oil from ower parts of C. nocturnum and its antibacterial property against the food-borne pathogenic bacteria. Therefore, the aims of the present study were (1) to examine the chemical composition of the essential oil of C. nocturnum L. by gas chromatography-mass spectrometry (GC-MS); and (2) to evaluate the antibacterial activity of essential oil from ower parts of C. nocturnum L. and various organic extracts (hexane, chloroform, ethyl acetate and methanol) against a diverse range of food-borne pathogenic bacteria with emphasis for the possible future use of the essential oil and plant extracts as an alternative to chemical bactericides for food preservation.
Materials and methods

type apparatus. The oil was dried over anhydrous Na2SO4 and preserved in a sealed vial at 4 C until further analysis.
Preparation of organic extracts

The air-dried ower parts (50 g) of C. nocturnum were extracted with hexane, chloroform, ethyl acetate and methanol separately at room temperature and the solvents were evaporated by vacuum rotary evaporator (EYELA N1000). The extraction process yielded in hexane (7.5 g), chloroform (6.6 g), ethyl acetate (5.4 g) and methanol (6.3 g) extracts. Solvents (analytical grade) for extraction were obtained from commercial sources (SigmaAldrich, St Louis, MO, USA).
Gas chromatography-mass spectrometry analysis

The GC-MS analysis of the essential oil was performed using a GC-MS (Model QP 2010, Shimadzu, Japan) equipped with a ZB-1 MS fused silica capillary column (30 m 0.25 i.d., lm thickness 0.25 lm). For GC-MS detection, an electron ionisation system with ionisation energy of 70 eV was used. Helium gas was used as a carrier gas at a constant ow rate of 1 mL min)1. Injector and mass transfer line temperature were set at 220 and 290 C, respectively. The oven temperature was programmed from 50 to 150 C at 3 C min)1, then held isothermal for 10 min and nally raised to 250 C at 10 C min)1. Diluted samples (1 100, v v, in methanol) of 1 lL was manually injected in the split less mode. The relative percentage of the oil constituents was expressed as percentage by peak area normalisation. The identity of the components of the essential oil was assigned by comparison of their retention indices, relative to a series n-alkane indices on the ZB-1 capillary column and GC-MS spectra from the Wiley 6.0 MS data and literature data (Adam, 2001).
Micro-organisms

Plant material

The owers of C. nocturnum were collected from Islamic University Campus, Kushtia, Bangladesh in December 2007. The taxonomic identication of plant materials was conrmed by a senior plant taxonomist Dr M. Oliur Rahman, Bangladesh National Herbarium, Dhaka, where a voucher specimen (DACB 32562) has been deposited.
Isolation of the essential oil

The air-dried ower parts (200 g) of C. nocturnum were subjected to hydrodistillation for 3 h using a Clevenger

The following food-borne pathogenic bacterial strains were used in the antibacterial test: S. aureus ATCC 6538, S. aureus KCTC 1916, L. monocytogenes ATCC 19166, L. monocytogenes ATCC 15313, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella typhimurium KCTC 2515, E. coli ATCC 8739, E. coli O157:H7 ATCC 43888, Enterobacter aerogenes KCTC 2190 and S. enteritidis KCTC 12021. The strains were obtained from the Korea Food and Drug Administration (KFDA), Daegu, Korea. Active cultures for experimental use were prepared by transferring a loopful of cells from stock cultures to asks and inoculated in Luria-Bertani (LB) broth medium at 37 C for 24 h. Cultures of each bacterial strains were maintained on LB agar medium at 4 C.

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Antibacterial activity assay

The dried extracts were dissolved in the same solvent used for their extraction to a nal concentration of 30 mg mL)1 and sterilised by ltration through 0.45 lm Millipore lters (Millipore Corp., Bedford, MA, USA). The antibacterial test was then carried out by agar disc diusion method (Murray et al., 1995) using 100 lL of standardised inoculum suspension containing 107 CFU mL)1 of bacteria. The essential oil was diluted 1:5 (v v) with methanol and aliquots of 5 lL were spotted onto the lter paper discs; while 10 lL of 30 mg mL)1 of each organic extract (300 lg per disc) was applied on the lter paper discs (6 mm diameter) and placed on the inoculated LB agar. Negative controls were prepared using the same solvents employed to dissolve the samples. Standard reference antibiotics, tetracycline and streptomycin (10 lg per disc, each from Sigma-Aldrich Co., St. Louis, MO, USA), were used as positive controls for the tested bacteria. The plates were incubated at 37 C for 24 h. Antibacterial activity was evaluated by measuring the diameter of the zones of inhibition against the tested bacteria. Where, 710 mm, weak inhibition; 1114 mm, moderate inhibition; >15 mm, strong inhibition. Each assay in this experiment was replicated three times.
Minimum inhibitory concentration

of L. monocytogenes ATCC19166, L. monocytogenes ATCC15313, S. aureus ATCC 6538 and P. aeruginosa KCTC 2004 in LB broth medium was inoculated with the MIC of the essential oil in 10 mL LB broth, and kept at 37 C (Bajpai et al., 2008). Samples for viable cell counts was taken out at 0, 20, 40, 60, 80 and 100 min time intervals. Enumeration of viable counts on LB plates were monitored as follows: after incubation, 1 mL of the resuspended culture was diluted into 9 mL buer peptone water, thereby diluting 10 fold. In total, 0.1 mL sample of each treatment was diluted and spread on the surface of LB agar. The colonies were counted after 24 h of incubation at 37 C. The controls were inoculated without essential oil for each bacterial strain with same experimental condition as mentioned above.
Statistical analysis

The essential oil and dierent organic extracts were assayed for antibacterial activity. Each experiment was run in triplicate, and mean values were calculated. A Students t-test was computed for the statistical significance of the results.
Results

Chemical composition of the essential oil

Minimum inhibitory concentration (MIC) of essential oil and extracts of hexane, chloroform, ethyl acetate and methanol, were tested by standard NCCL method (NCCLS, 2000). Active cultures for MIC determination were prepared by transforming a loopful of cells from the stock cultures to asks and inoculated in LB medium and incubated at 37 C for 24 h. The cultures were diluted with LB broth to achieve optical density of 107 CFU mL)1 for the test organisms at 600 nm by UV Vis Spectrophotometer Optizen 2120UV & Optizen III (Shin et al., 2007). Dilutions, to get the nal concentration ranging from 0 to 1000 lg mL)1 of essential oil and extracts of hexane, chloroform, ethyl acetate and methanol in LB broth medium were prepared in a 96-well microplates. Finally, 20 lL inoculum of each bacteria strain (107 CFU mL)1) was inoculated on to the microplates and the tests were performed in a volume of 200 lL. The plates were incubated at 37 C for 24 h. The lowest concentration of the test samples, which did not show any visual growth of tested organisms after macroscopic evaluation, was determined as MIC, which was expressed in lg mL)1.
Effect of essential oil on viable counts of bacteria

The hydrodistillation of the air-dried owers of C. nocturnum gave the dark yellowish oil with a yield of 0.34% (w w). Gas chromatography-mass spectrometry analyses of the oil led to the identication of 47 dierent compounds, representing 93.28% of the total oil. The identied compounds are listed in Table 1 according to their elution order on a ZB-1 capillary column. The major compounds detected were phenylethyl alcohol (27.45%), benzyl alcohol (12.21%), eicosane (5.62%), eugenol (5.59%), n-tetracosane (4.42%), caryophyllene oxide (3.15%), 1-hexadecanol (2.75%), methoxyeugenol (2.45%) and benzaldehyde (2.32%). Hexadecanoic acid (1.71%), 1-nonadecanol (1.65%), heneicosane (1.55%), methyl anthranilate (1.44%), nonadecene (1.37%), nerolidol (1.31%), tetradecanal (1.28%) and citronellal (1.23%) were also found to be the minor components of C. nocturnum oil in the present study.
Antibacterial activity

For viable counts, each of the tubes containing bacterial suspension (approximately 107 CFU mL)1)

The in vitro antibacterial activity of essential oil and various extracts (hexane, chloroform, ethyl acetate and methanol) of C. nocturnum against the employed bacteria was qualitatively assessed by the presence or absence of inhibition zones. According to the results given in Table 2, a total of eleven food-borne pathogenic bacteria, including ve Gram-positive and six Gram-negative bacteria were tested. The oil exhibited

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Table 1 Chemical composition of the essential oil from ower parts

Table 1 (Continued) Compound RIa Composition (%)

Cestrum nocturnum L
Compound Hydrocarbons 8-Methyl-1-unde cene n-Hexadecane Nonadecene Eicosane He ne ico sane n-Tetracosane Aldehydes and ketones Benzaldehyde Geranyl acetone Benzofuranone Dicyclohexyl ketone Tetrade canal Pentade canal He xade canal Octade canal Alcohols Benzyl alcohol Phenyl ethyl alcohol Cycloheptanol n-Undecanol Dodecanol Pentadecanol 1-Hexadecanol Heptadecanol E.E-2, 1 3-Octadecadien-1-ol 1-Nonadecanol Phenols Durenol Eugenol Methoxyeugenol Fatty adds Tetradecanoic acid Hexadecanoic acid Pentadecanoic acid (Z)-9-Octadecenoic acid 9,12-Octadecadienoic acid Eicosanoic acid Esters Vinyl cyclohexanecarboxylate Methyl anthranilate Famesyl acetate Monoterpene Azulene Oxygenated monoterpene p-Menth-8-en-2-ol Sesquiterpenes Famesene Caiyophyllene Oxygenated sesquiterpenes Caiyophyllene oxide Nerolidol Phytol Nitrogenous compound Indole RI
a

Composition (%)

1140 1612 1918 2009 2109 2407 982 1420 1426 1576 1601 1701 1800 1999 1036 1136 1306 1357 1457 1755 1854 1954 2069 2153 1354 1392 1581 1769 1968 2101 2175 2183 2366 1137 1372 1834 1386 1196 1320 1494 1567 1564 2045 1174

0.38 0.41 1.37 5.62 1.55 4.42 2.32 0.12 0.32 0.45 1.28 1.13 0.47 0.91 12.21 27.45 0.48 0.44 0.44 0.32 2.75 0.62 0.34 1.65 0.45 5.59 2.45 0.89 1.71 0.23 0.37 0.65 1.23 0.35 1.44 0.32 0.33 0.36 0.79 0.68 3.15 1.31 0.93 0.43

Monoterpenoid Citronellal Others trans-Z-d-Bisabolene ep oxide 2, 5-Dim etho xy ac et anili de

1125 1531 1671 Total

1.23 0.75 0.19 93.28

Retention index relative to n-alkanes on ZB-1 capillary column.

antibacterial activity against all ve Gram-positive and three Gram-negative bacteria at the concentration of 5 lL of 1:5 (v v) dilution with methanol. The oil exhibited a potent inhibitory eect against S. aureus (ATCC 6538 and KCTC 1916), L. monocytogenes (ATCC 19166 and ATCC 15313), B. subtilis ATCC 6633, P. aeruginosa KCTC 2004, S. typhimurium KCTC 2515 and E. coli ATCC 8739 with diameter of inhibition zones ranging from of 9.818.2 mm, as shown in Table 2. Various organic extracts of C. nocturnum also revealed a great potential of antibacterial activity against all ve Gram-positive and three Gram-negative bacteria (P. aeruginosa KCTC 2004, S. typhimurium KCTC 2515 and E. coli ATCC 8739), at the concentration of 300 lg per disc (Table 2). Methanol extract showed the strongest antibacterial eect against S. aureus (ATCC 6538 and KCTC 1916), L. monocytogenes ATCC 19166 and B. subtilis ATCC 6633 with their respective diameter zones of inhibition of 18.1, 18.2, 17.0 and 16.4 mm, as compared with standard drug streptomycin. On the other hand, hexane, chloroform and ethyl acetate extracts showed interesting antibacterial eect with inhibition zones in the range of 10.113.2, 10.114.1 and 11.116.2 mm, respectively. In this study, in some cases, the oil and organic extracts (chloroform, ethyl acetate and methanol) exhibited higher antibacterial activity compared with streptomycin, while tetracycline showed higher activity in some other cases than the essential oil and solvent extracts. The blind control did not inhibit the growth of the tested bacteria. The oil and various extracts (hexane, chloroform, and ethyl acetate) from C. nocturnum exhibited a moderate inhibitory eect against P. aeruginosa KCTC 2004, S. typhimurium KCTC 2515 and E. coli ATCC 8739, with diameter zones of inhibition in the range of 9.813.3 and 10.113.2 mm, respectively. No inhibitory eect was observed against E. coli O157:H7 ATCC 43888, E. aerogenes KCTC 2190 and S. enteritidis KCTC 12021 in all cases.
Minimum inhibitory concentration

As shown in Table 3, the MIC values for the oil were found lower for L. monocytogenes (ATCC 19166 and

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Table 2 Antibacterial activity of essential oil and various extracts of Cestrum nocturnum L. against food-borne pathogenic and spoilage bacteria Zones of inhibition (mm) Various extractsb Micro-organism S. aureus ATCC 6538 S. aureus KCTC 1916 L. monocytogenes ATCC 19166 L. monocytogenes ATCC 15313 B. subtilis ATCC 6633 P. aeruginosa KCTC 2004 S. typhimurium KCTC 2515 E. coli ATCC 8739 E. coli 0157H7ATCC43888 E. aerogenes KCTC 2190 S. enteritidis KCTC 12021 Essential oila 18.2 1.1 16.2 1.5 16.3 0.9 14.1 1.1 15.3 0.7 13.3 1.2 9.8 1.5 10.0 0.6 nd nd nd Hexane 13.1 13.2 12.1 12.5 13.2 11.6 10.1 nd nd nd nd 0.7 0.5 1.1 0.7 0.8 1.1 1.1 CHCl3 14.0 13.6 14.1 14. 1 12.3 11. 0 10.2 10.1 nd nd nd 1.6 1.2 0.5 1.1 1.2 1.2 1.2 1.2 EtOAc 16.2 15.6 15.2 14.1 13.6 13.2 12.3 11.1 nd nd nd 1.5 1.2 1.0 1.2 0.6 1.1 1.6 0.7 MeOH 18.1 18.2 17.0 16.7 16.4 15.1 13.2 11. 8 nd nd nd 1.4 1.2 1.2 1.2 0.7 1.4 1.1 1.7 Antibioticsc TC 17.8 18. 1 17.2 18.4 18.3 17.9 18.0 153 16.3 15 .1 18.3 0.6 0.6 0.7 0.5 0.5 1.2 0.6 1.2 0.5 1.0 1.3 SM 13.2 13. 1 12.6 16.2 14.3 18.0 13.3 14.0 12.3 10.0 11.3 0.6 0.5 0.7 1.2 0.6 0.5 0.6 0.7 1.1 0.5 0.6

Values are given as mean SD (n = 3). nd, not detected. a Diameter of inhibition zones of essential oil including diameter of disc 6 mm (tested at a volume of 5 lL per disc). b Various extract (300 lg disc) Where, 710 mm, weak inhibition, 1114 mm, moderate inhibition, >15 mm, strong inhibition. c Standard antibiotics: TC, tetracyclme and S.M, streptomycin (10 lg per disc).

Table 3 Minimum inhibitory concentration of essential oil and various

extracts of Cesturm nocturnum L. against food-borne pathogenic and spoilage bacteria

Minimum inhibitory concentration (MIC)a Various extracts Micro-organism S. aureus ATCC 6538 S. aureus KCTC 1916 L. monocytogenes ATCC 19166 L. monocytogenes ATCC 15313 B. subtils ATCC 6633 P. aeruginosa KCTC 2004 S. typhimurium KCTC 2515 E. coli ATCC 8739 E. coli 0157H7ATCC43888 E aerogenes KCTC 2190 S enteritidis KCTC 12021 Essential oil Hexane CHCl3 EtOAc MeOH 12.5 25.0 62.5 12.5 25.0 12.5 25.0 50.0 nd nd nd 25.0 25.0 25.0 50.0 25.0 50.0 50.0 nd nd nd nd 12.5 25.0 12.5 25.0 25.0 25.0 50.0 50.0 nd nd nd 62.5 25.0 62.5 12.5 12.5 25.0 25.0 50.0 nd nd nd 62.5 12.5 62.5 12.5 12.5 25.0 50.0 50.0 nd nd nd

antibacterial activity by MICs than hexane and chloroform extracts. In this study, the Gram-positive bacteria were found to be more susceptible to the essential oil and various solvent extracts than did Gram-negative bacteria.
Effect of essential oil on viable counts of bacteria

nd, not detected. a Minimum inhibitory concentration (values in lg mL)1).

ATCC 15313) S. aureus ATCC 6538 and P. aeruginosa KCTC 2004 (62.5125 lg mL)1) than for S. aureus KCTC 1916, B. subtilis ATCC 6633, S. typhimurium KCTC 2515 and E. coli ATCC 8739 (250500 lg mL)1). On the other hand, MIC values of various solvent extracts against the tested bacteria were found in the range of 62.5500 lg mL)1 (Table 3). Methanol and ethyl acetate extracts showed higher

Based on the susceptibility, further, elaborative study carried out on L. monocytogenes ATCC19166, L. monocytogenes ATCC15313, S. aureus ATCC 6538 and P. aeruginosa KCTC 2004, displayed dierent sensitivities of the essential oil. The eects of essential oil on the growth of all the tested bacterial strains demonstrated the reduced viability of the tested bacteria at MIC concentration of the essential oil. Complete inhibition of both strains of L. monocytogenes ATCC19166 and L. monocytogenes ATCC15313 was observed at MIC concentration of the essential oil at 20 and 40 min exposure, respectively. Also the steep decline in CFU numbers was observed at 60 min exposure against S. aureus ATCC 6538 and P. aeruginosa KCTC 2004. Exposure of 80 min of the essential oil MIC concentration revealed complete inhibition of CFU numbers against all the bacterial strains tested (Fig. 1).
Discussion

Plant based secondary metabolites such as essential oil and extracts are widely used in the food industry and are considered generally recognised as safe (GRAS). They usually contain more than a single compound with

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Figure 1 Effect of Cestrum nocturnum L. essential oil (minimum inhibitory concentration) on viability of the tested bacteria. CT, control without treatment.

antimicrobial activity. Hence, using essential oil extracts has, as a consequence, to take advantage of all active compounds present in essential oil extracts (Hao et al., 1998). Plants-derived essential oils because of their antimicrobial content possess potential signicance as naturally occurring agents for food preservation. Many volatile compounds naturally occurring in various essential oils possess strong antibacterial activities, thereby considering as natural antibacterial agents to inhibit the growth of food-borne pathogens (Cowan, 1999). The renewal of interest in food industry, and increasing consumer demand for eective natural products means that quantitative data on plant based essential oils is required. Various publications documented the antibacterial activity of essential oil constituents and plant extracts. In recent years, several researchers have also reported that the mono- and sesquiterpenoids are the major components of essential oils which exhibit potential antibacterial activities (Shunying et al., 2005). Also, the results of the antibacterial screening showed that essential oil and various extracts of C. nocturnum have potential activity against some of the bacterial strains such as S. aureus (ATCC 6538 and KCTC 1916), L. monocytogenes (ATCC 19166 and ATCC 15313), B. subtilis ATCC 6633, P. aeruginosa KCTC 2004, S. typhimurium KCTC 2515 and E. coli ATCC 8739. This activity could be attributed to the presence of oxygenated mono- and sesquiterpene hydrocarbons, and these nding are in agreement with the previous reports (Larsen & Knochel, 1997). C. nocturnum mediated oil also contained high percentage of phenylethyl alcohol, benzyl alcohol, eicosane, eugenol, n-tetracosane, caryo-

phyllene oxide, 1-hexadecanol, methoxyeugenol and benzaldehyde, as earlier reported the major components of the various essential oils, which have potential antibacterial properties (El-Sakhawy et al., 1998; Thangadurai et al., 2002; Deba et al., 2008). Those claims are further supported by our ndings; indicating high contents of phenylethyl alcohol, benzyl alcohol, eicosane, eugenol, n-tetracosane, caryophyllene oxide, 1-hexadecanol, methoxyeugenol and benzaldehyde; comprising 65.96% of the oil (Table 1). The antibacterial activity of individual component of essential oils such as phenylethyl alcohol, benzyl alcohol or eugenol has been reported previously (Lucchini et al., 1990; Jirovetz et al., 2006). On the other hand, the components in lower amount such as hexadecanoic acid, 1-nonadecanol, heneicosane, methyl anthranilate, nonadecene, nerolidol, tetradecanal and citronellal also contributed to antimicrobial activity of the oil (Jeongmok et al., 1995; El-Sakhawy et al., 1998; Khan et al., 2002; Deba et al., 2008). It is also possible that the minor components might be involved in some type of synergism with the other active compounds (Marino et al., 2001). Also, the results from viable count assay revealed that exposure of the MIC concentration of the oil had a severe eect on the cell viability of the tested bacteria. L. monocytogenes ATCC19166 and L. monocytogenes ATCC15313 were found to be more sensitive to the oil. The oil also exerted its maximum bacterial activity against S. aureus ATCC 6538 and P. aeruginosa KCTC 2004, as evident by the signicant reduction in microbial counts at 60 min exposure and complete inhibition of cell viability at 80 min exposure of essential oil. Deans et al. (1995) investigated the susceptibility of Gram-positive and Gram-negative bacteria to plant volatile oils and found no evidence for a dierence in sensitivity between Gram-negative and Gram-positive organisms. However, some oils appeared more specic, exerting a greater inhibitory activity against Grampositive bacteria. It is often reported that Gram-negative bacteria are more resistant to the plant-based essential oils. The hydrophilic cell wall structure of Gramnegative bacteria is constituted essentially of a lipopolysaccharide (LPS) that blocks the penetration of hydrophobic oil and avoids the accumulation of essential oils in target cell membrane (Bezic et al., 2003). This is the reason that Gram-positive bacteria were found to be more sensitive to the essential oil and various extracts of C. nocturnum than those of Gram-negative bacteria. In this study, we found that essential oil and various extracts from C. nocturnum growing in Bangladesh inhibited the growth of some representative food-borne pathogenic bacteria. Therefore, essential oils and plant extracts are being considered as potential alternatives to synthetic bactericides or as leading compounds for new classes of natural bactericides.

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In conclusion, the results of our study suggest the possibility of using the extract or oil of C. nocturnum as natural antimicrobials in food industry to control foodborne pathogens. The use of plant extracts and essential oils in consumer goods is expected to increase in the future because of the risk of green consumerism, which stimulates the use and development of products derived from plants, as both consumers and regulatory agencies are more comfortable with the use of natural antimicrobials. It was found that the essential oils of Salvia ocinalis L. and Schinus molle L. had preservative eect against Salmonella inoculated in minced beef meat. Such ndings would also be conrmed in further studies to establish the real application of C. nocturnum essential oil or extracts in foods.
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International Journal of Food Science and Technology 2009

2009 The Authors. Journal compilation 2009 Institute of Food Science and Technology