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Anti-listerial activity of plant essential oils from western region of Argentina

Anti-listerial activity of plant essential oils from western region of Argentina Annals of Microbiology, 56 (4) 369-371 (2006) 1 1 2 2 2 M. Laura VACA RUIZ *, Analía LACIAR , Osvaldo J. DONADEL ; J. Roberto SAAD , Roberto CARRIZO FLORES 1 2 Área Microbiología, Área de Química Orgánica INTEQUI-CONICET, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, Chacabuco y Pedernera, San Luis, 5700, Argentina Received 30 June 2006 / Accepted 16 October 2006 Abstract - Listeria monocytogenes is an important foodborne pathogen due to the severity of infection with a high mortality rate. In the past few years, there has been an increase in the use of naturally derived compounds such as plant extracts or essential oils as antimicrobials in food. The objective of the present study was to determine the antimicrobial effect of the Flourensia oolepis, Baccha- ris salicifolia, and Artemisia echegarayi essential oils on fourteen strains of L. monocytogenes, by the disk diffusion method. The results indicate that the essential oils tested could potentially be used to inhibit L. monocytogenes, but appropriate applications in food should be validated. Key words: Baccharis salicifolia, Flourensia oolepis, Artemisia echegarayi, essential oils, Listeria monocytogenes, antibacterial activity. INTRODUCTION pounds such as plant extracts or essential oils as preserva- tive agents in food (Chorianopoulos et al., 2004). In this con- Listeria monocytogenes is an important foodborne pathogen text, essential oils derived from plants of Flourensia oolepis, which is the cause of listeriosis, one of the most deadly bac- Baccharis salicifolia, and Artemisia echegarayi, all of which terial infections currently known, with a mean mortality rate grow in the western region of Argentina (Mendoza, San Luis, in humans of 20 to 30% or higher despite early antibiotic and San Juan), have been studied for this purpose. Flouren- treatment (Vazquez et al., 2001). Contamination of food with sia oolepis has been shown to have insecticidal activity (Gar- L. monocytogenes is almost inevitable due to its ubiquitous cia et al., unpublished results). The genus Baccharis has been nature in the environment (Smith-Palmer et al., 2001). In found to possess diuretic, digestive and liver protector bioac- addition, L. monocytogenes exhibits a number of charac- tivities, among others (Cobos et al., 2001). Antibacterial teristics that make its presence in food processing environ- activity has been found in Baccharis grisebachii (Feresin et ments undesirable as well as inevitable: it can grow at refrig- al., 2003) and B. salicifolia (Carrizo Flores et al., unpublished eration temperatures, it has the ability to survive in acid con- results), probably caused by the presence of high concen- ditions and resist up to 10% sodium chloride solution (Laciar tration of terpene hydrocarbons. In addition, the Artemisia et al., 2002), and it can survive in biofilms on equipment in genus is known for possessing a wide spectrum of activities. food processing plants (Lundén et al., 2000). On the other For example Artemisia douglassiana is used as gastric cyto- hand, L. monocytogenes can contaminate products mainly protector and topic bactericidal on skin (Giordano et al., after thermal treatments, and thus, ready-to-eat foods that 1992; Setzer et al., 2004). Artemisia annua has antimalar- are commonly consumed without further cooking are of par- ial, anti-inflammatory, antitumor and allelopathic activities ticular concern (Kathariou, 2002). (Juteau et al., 2002). Furthermore, we have demonstrated Effective methods for killing L. monocytogenes in food the antibacterial activity of A. echegarayi (Carrizo et al., may reduce the likelihood of foodborne listeriosis outbreaks unpublished results). and reduce economic losses to the food industry (Nair et al., The objective of the present study was to determine the 2005). A wide variety of different chemical and synthetic inhibitory capacity of F. oolepis, B. salicifolia and A. compounds have been used as antimicrobial agents to inhib- echegarayi essential oils on L. monocytogenes strains, for it L. monocytogenes in food. However, in the past few years, potential application as preservatives in food industries. due to concerns regarding the safety of synthetic antimi- crobial substances, there has been an increase in consumer demand for naturally processed food (Hao et al., 1998). MATERIALS AND METHODS This has resulted in greater use of naturally derived com- Plant material. Aerial parts of B. salicifolia and F. oolepis were collected in February 2001, at El Volcan, Province of San Luis, Argentina and A. echegarayi was collected in March * Corresponding author. Phone: +54-02652-423789; Fax: +54-02652-422282; E-mail address: mlvaca@unsl.edu.ar 2005 at Potrerillos, Province of Mendoza, Argentina. A vouch- 370 M.L. Vaca Ruiz et al. er of each sample was deposited at the Herbario of the Uni- plates were allowed to dry for 5 min at room temperature. versidad Nacional de San Luis under numbers: 492- Del Sterilised paper disks (Britania) of 6 mm diameter were Vitto [A. echegarayi Hieron. (Asteraceae)], 8487-Del Vitto used. Ten microlitres of each essential oil and vegetable oil [F. oolepis Blake (Asteraceae)] and 9186-Del Vitto [B. sali- (negative control) were added to impregnate paper disks and cifolia (Ruiz & Pavon) Pers]. allowed dry for 15 min. Commercial gentamicin disks (10 µg, Britania) were used as positive control. The disks were then Extraction of essential oil. Fresh aerial parts (5000 g) of placed aseptically over the surface of the bacterial cultures each plant were cut into small pieces and subjected to on MH plates and incubated at 37 C for 24 h. After incuba- steam-distillation at 96 C for 3 h using a Clevenger type tion, the inhibition zones around paper disks were measured -1 apparatus. The oils obtained (0.56 g kg ) were dried over accurately using a metric ruler. Assays were carried out on anhydrous sodium sulphate (Deans and Svoboda, 1990). duplicate MH plates for each L. monocytogenes strain. The experiments were replicated at least twice and the data Cultures. Fourteen strains of L. monocytogenes, listed in were analysed using InfoStat program to calculate the vari- Table 1, were used for the study. This number includes three ance analysis. Significant differences in the antibacterial strains from the American Type Culture Collection and one effects between the essential oils studied were determined strain from the Pasteur Institute Collection. The remaining by Duncan test (P < 0.01). L. monocytogenes strains were isolated in our laboratory from human, animal, and food samples. Each strain of L. monocytogenes was individually cultured RESULTS AND DISCUSSION in Trypticase Soy broth (Britania, Argentina) at 37 C for 24 h. Following incubation, the bacterial cells were separated by The antibacterial effect of three essential oils on different L. centrifugation (5000 rpm for 15 min at 4 C), washed twice monocytogenes strains is shown in Table 1. The average inhi- with sterile phosphate buffered saline (PBS, pH 7.2), and bition zone of F. oolepis against L. monocytogenes ranged finally resuspended in PBS. The cultures were appropriate- from 18.7 ± 2.6 mm (L. monocytogenes CLIP 74903) to 8.7 ly diluted in PBS to obtain a concentration of 10 CFU/ml, (0.5 ± 0.9 mm (L. monocytogenes 4425), yielding a mean on Mac Farland scale). The bacterial population in each cul- inhibitory zone of 12.64 mm on L. monocytogenes strains. ture was verified by spreading 100 ml aliquots of the appro- The average inhibition zone of B. salicifolia ranged from priately diluted cultures on Trypticase Soy agar (Britania) 12.2 ±1.8 mm (L. monocytogenes CLIP 74903) to 8.0 ±0.7 plates, with incubation at 37 C for 24 h. mm (L. monocytogenes 4425), yielding a mean inhibitory zone of 10.63 mm on L. monocytogenes strains. Artemisia Determination of antibacterial effect. The antibacterial echegarayi essential oil showed the same inhibition pattern: effect of F. oolepis, B. salicifolia, and A. echegarayi on the 12.5 ± 1.1 mm (L. monocytogenes CLIP 74903) to 7.7 ± 1.0 fourteen strains of L. monocytogenes was determined by the mm (L. monocytogenes 4425), yielding a mean inhibitory standard disk diffusion technique (CLSI, 2005). A population zone of 9.56 mm on L. monocytogenes strains. Gentamicin of approximately 10 CFU/ml of each L. monocytogenes resulted in inhibition zones ranging from 33.5 ± 1.9 mm (L. strain was inoculated on duplicate plates containing Müeller monocytogenes I) to 23.8 ± 1.8 mm (L. monocytogenes Hinton agar (MH, Britania) using sterile cotton swabs. The 4425). Vegetable oil produced no L. monocytogenes inhibi- TABLE 1 – Inhibitory effect of Flourensia oolepis, Baccharis salicifolia, Artemisia echegarayi essential oils on Listeria monocytogenes strains a b c d e L. monocytogenes Source/origin GN (mm)* VO (mm) Fo (mm)* Bs (mm)** Ae (mm)** strains Mean ± SE Mean ± SE Mean ± SE Mean ± SE Mean ± SE A Human 30.0 ± 2.5 0 14.0 ± 2.9 10.4 ± 1.1 9.25 ± 0.6 D Animal 31.7 ± 2.0 0 11.5 ± 1.9 11.0 ± 1.4 8.50 ± 1.5 E Animal 30.0 ± 1.4 0 12.0 ± 1.8 10.5 ± 1.0 9.50 ± 0.6 G Food 29.4 ± 0.5 0 10.5 ± 2.3 9.00 ± 0.8 8.00 ± 1.0 H Food 29.2 ± 1.6 0 13.0 ± 1.4 10.5 ± 1.7 8.00 ± 1.1 I Food 33.5 ± 1.9 0 14.2 ± 3.7 12.0 ± 1.4 10.3 ± 0.7 K Food 30.2 ± 1.3 0 11.5 ± 1.0 10.2 ± 0.8 9.25 ± 0.6 MF2 Human 25.4 ± 2.3 0 9.70 ± 2.2 11.0 ± 1.0 10.0 ± 1.5 56 Food 29.2 ± 1.9 0 11.5 ± 2.3 9.40 ± 0.5 10.0 ± 0.6 4425 Human 23.8 ± 1.8 0 8.70 ± 0.9 8.00 ± 0.7 7.70 ± 1.0 35152 ATCC 30.2 ± 1.7 0 12.5 ± 0.7 11.3 ± 0.6 9.75 ± 0.0 74903 CLIP 31.8 ± 2.4 0 18.7 ± 2.6 12.2 ± 1.8 12.5 ± 1.1 19112 ATCC 30.2 ± 2.7 0 15.3 ± 0.6 11.2 ± 1.5 10.0 ± 2.1 19115 ATCC 29.5 ± 1.5 0 13.6 ± 0.6 12.0 ± 1.7 11.0 ± 2.8 a b c d e f GN: gentamicin, VO: vegetable oil, Fo: Fluorensia oolepis, Bs: Baccharis salicifolia, Ae: Artemisia echegarayi, ATCC: American Type Culture Collection, CLIP: Listeria Collection of the Pasteur Institute. * Very significative (P < 0.01), ** Significative (P < 0.05). Ann. Microbiol., 56 (4), 369-371 (2006) 371 tion zone, indicating that it does not have any antibacterial ty testing. Fifteenth Informational Supplement. Disk diffu- sion. M100-S15. Vol. 25 No. 1. effect on this pathogen. Although all strains of L. monocytogenes varied in their Cobos M., Rodriguez J., Oliva M., Demo M., Faillaci S., Zygadlo J. (2001). Composition and antimicrobial activity of the essen- antimicrobial susceptibility, F. oolepis exhibited the highest tial oil of Baccharis notosergila. Planta Med., 67: 84-86. activity with a highly significant difference as compared with Cosentino S., Tuberoso C., Pisano B., Satta M., Mascia V., Arzedi the other oils (P < 0.01). E., Palmas F. (1999). In vitro antimicrobial activity and chem- Gentamicin was used as a positive control in the study ical composition of Sardinian Thymus essential oils. Lett. Appl. because L. monocytogenes has been reported to be highly Microbiol., 29: 130-135. sensitive to this antibiotic. In fact, gentamicin and ampicillin Deans S., Svoboda K. (1990). The antimicrobial properties of are usually recommended for the treatment of listeriosis marjoram (Origanum majorana L.) volatile oil. Flavour Frag. (Hof et al., 1997). In a study on black seed (Nigella sativa) J., 5: 187-190. oil Nair et al. (2005) found that gentamicin inhibition zones Feresin G., Tapia A., Gimenez A., Gutierrez Ravelo A., Zacchino S., were smaller than those of black seed oil. The high activity Sortino M., Schmeda-Hirschman G. (2003). Constituents of the Argentinian medicinal plant Baccharis grisebachii and their of black seed oil is due to presence of carvacrol, thymol and antimicrobial activity. J. Ethnopharmacol., 89: 73-80. thymohydroquinone, which are well known for their anti- bacterial activities. In our study, gentamicin showed inhibi- Giordano O., Pestchanker M., Guerreiro E., Saad J.R., Enriz R., Rodriguez A., Jauregui E., Guzman J., Maria A., Wendel G. tion zones wider than those of essential oils. The moderate (1992). Structure-activity relationship in the gastric cytopro- antilisterial activity of F. oolepis, A. echegarayi, and B. sali- tective effect of several sesquiterpene lactones. J. Med. Chem., cifolia essentials oils can be explained by their chemical 35: 2452-2458. composition. Although they do not present phenolic com- Hao Y., Brackett R.E., Doyle M.P. (1998). Inhibition of Listeria pounds, their antibacterial activity is probably due, at least monocytogenes and Aeromonas hydrophila by plants extracts in part, to the in situ formation of active compounds from in refrigerated cooked beef. J. Food Prot., 61: 307-312. their precursors, e.g. γ-terpinene (1-methyl-4-(1- Hof H., Nichterlein T., Kretschmar M. (1997). Management of lis- methylethyl)-1,4-cyclohexadiene) is the precursor of thymol teriosis. Clin. Microbiol. Rev., 10: 345-357. (5-methyl-2-(1-methylethyl) phenol) (Cosentino et al., Juteau F., Massoti V., Bessiere J., Dherbomez M., Viano J. (2002). 1999). The essential oils evaluated in this work have a great Antibacterial and antioxidant activities of Artemisia annua variety of phytochemicals that could be considered respon- essential oil. Fitoterapia, 73: 532-535. sible for a larger or smaller part of the antibacterial activity. Kathariou S. (2002). Listeria monocytogenes virulence and patho- Although they usually occur as complex mixtures, their activ- genecity, a food safety perspective. J. Food Prot., 65: 1811- ity can be accounted for in terms of their major components. Research into the antimicrobial actions of monoterpenoids Laciar A.L., Centorbi O.N.P. de (2002). Listeria species in seafood: isolation and characterization of Listeria spp. from seafood in suggests that they diffuse into and damage cell membrane San Luis, Argentina. Food Microbiol., 19: 645-651. structures (Sikkema et al., 1995). Even though the exact Lundén J.M., Miettinen M.K., Autio T.J., Korkeala H.J. (2000). Per- nature of the antibacterial activity is not known, our results sistent Listeria monocytogenes strains show enhanced adher- indicate that the essential oils tested possess moderate ence to food contact surface after short contact times. J. Food activity against L. monocytogenes. However, appropriate Prot., 63: 1204-1207. applications in food should be validated. Nair M.K., Vasudevan P., Venkitanarayanan K. (2005). Antibacte- rial effect of black seed oil on Listeria monocytogenes. Food Acknowledgements Control, 16: 395-398. We have to thank to UNSL (Projects 7301 and 8802), CON- Setzer W., Vogler B., Schmidt J., Leahy J., Rives R. (2004). Antimi- ICET (PIP 6228) for the financial support of this study. crobial activity of four Artemisia species of Iran. Fitoterapia, 75: 192-200. Sikkema J., de Bont J. A., Poolman B. (1995). Mechanism of mem- REFERENCES brane toxicity of hydrocarbons. Microbiol. Rev., 59: 201-222. Smith-Palmer A., Stewart J., Fyfe L. (2001). The potential appli- Chorianopoulos N., Kalpoutzakis E., Aligiannis N., Mitaku S., Nichas cation of plant essential oils as natural food preservatives in G.J., Haroutounian S.A. (2004). Essential oils of Satureja, Origanum, and Thymus species: chemical composition and soft cheese. Food Microbiol., 18: 463-470. antibacterial activities against foodborne pathogens. J. Agric. Vazquez Boland J.A., Kuhn M., Berche P., Chakraborty T., Food Chem., 52: 8261-8267. Dominguez-Bernal G., Goebel W., Gonzalez- Zorn B., Wehland CLSI-Clinical and Laboratory Standards Institute (formely NCCLS) J., Kreft J. (2001). Listeria pathogenesis and molecular viru- (2005). Performance standards for antimicrobial susceptibili- lence determinants. Clin. Microbiol. Rev., 14: 584-640. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Annals of Microbiology Springer Journals

Anti-listerial activity of plant essential oils from western region of Argentina

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Springer Journals
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Copyright © 2006 by University of Milan and Springer
Subject
Life Sciences; Microbiology; Microbial Genetics and Genomics; Microbial Ecology; Fungus Genetics; Medical Microbiology; Applied Microbiology
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10.1007/BF03175034
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Abstract

Annals of Microbiology, 56 (4) 369-371 (2006) 1 1 2 2 2 M. Laura VACA RUIZ *, Analía LACIAR , Osvaldo J. DONADEL ; J. Roberto SAAD , Roberto CARRIZO FLORES 1 2 Área Microbiología, Área de Química Orgánica INTEQUI-CONICET, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, Chacabuco y Pedernera, San Luis, 5700, Argentina Received 30 June 2006 / Accepted 16 October 2006 Abstract - Listeria monocytogenes is an important foodborne pathogen due to the severity of infection with a high mortality rate. In the past few years, there has been an increase in the use of naturally derived compounds such as plant extracts or essential oils as antimicrobials in food. The objective of the present study was to determine the antimicrobial effect of the Flourensia oolepis, Baccha- ris salicifolia, and Artemisia echegarayi essential oils on fourteen strains of L. monocytogenes, by the disk diffusion method. The results indicate that the essential oils tested could potentially be used to inhibit L. monocytogenes, but appropriate applications in food should be validated. Key words: Baccharis salicifolia, Flourensia oolepis, Artemisia echegarayi, essential oils, Listeria monocytogenes, antibacterial activity. INTRODUCTION pounds such as plant extracts or essential oils as preserva- tive agents in food (Chorianopoulos et al., 2004). In this con- Listeria monocytogenes is an important foodborne pathogen text, essential oils derived from plants of Flourensia oolepis, which is the cause of listeriosis, one of the most deadly bac- Baccharis salicifolia, and Artemisia echegarayi, all of which terial infections currently known, with a mean mortality rate grow in the western region of Argentina (Mendoza, San Luis, in humans of 20 to 30% or higher despite early antibiotic and San Juan), have been studied for this purpose. Flouren- treatment (Vazquez et al., 2001). Contamination of food with sia oolepis has been shown to have insecticidal activity (Gar- L. monocytogenes is almost inevitable due to its ubiquitous cia et al., unpublished results). The genus Baccharis has been nature in the environment (Smith-Palmer et al., 2001). In found to possess diuretic, digestive and liver protector bioac- addition, L. monocytogenes exhibits a number of charac- tivities, among others (Cobos et al., 2001). Antibacterial teristics that make its presence in food processing environ- activity has been found in Baccharis grisebachii (Feresin et ments undesirable as well as inevitable: it can grow at refrig- al., 2003) and B. salicifolia (Carrizo Flores et al., unpublished eration temperatures, it has the ability to survive in acid con- results), probably caused by the presence of high concen- ditions and resist up to 10% sodium chloride solution (Laciar tration of terpene hydrocarbons. In addition, the Artemisia et al., 2002), and it can survive in biofilms on equipment in genus is known for possessing a wide spectrum of activities. food processing plants (Lundén et al., 2000). On the other For example Artemisia douglassiana is used as gastric cyto- hand, L. monocytogenes can contaminate products mainly protector and topic bactericidal on skin (Giordano et al., after thermal treatments, and thus, ready-to-eat foods that 1992; Setzer et al., 2004). Artemisia annua has antimalar- are commonly consumed without further cooking are of par- ial, anti-inflammatory, antitumor and allelopathic activities ticular concern (Kathariou, 2002). (Juteau et al., 2002). Furthermore, we have demonstrated Effective methods for killing L. monocytogenes in food the antibacterial activity of A. echegarayi (Carrizo et al., may reduce the likelihood of foodborne listeriosis outbreaks unpublished results). and reduce economic losses to the food industry (Nair et al., The objective of the present study was to determine the 2005). A wide variety of different chemical and synthetic inhibitory capacity of F. oolepis, B. salicifolia and A. compounds have been used as antimicrobial agents to inhib- echegarayi essential oils on L. monocytogenes strains, for it L. monocytogenes in food. However, in the past few years, potential application as preservatives in food industries. due to concerns regarding the safety of synthetic antimi- crobial substances, there has been an increase in consumer demand for naturally processed food (Hao et al., 1998). MATERIALS AND METHODS This has resulted in greater use of naturally derived com- Plant material. Aerial parts of B. salicifolia and F. oolepis were collected in February 2001, at El Volcan, Province of San Luis, Argentina and A. echegarayi was collected in March * Corresponding author. Phone: +54-02652-423789; Fax: +54-02652-422282; E-mail address: mlvaca@unsl.edu.ar 2005 at Potrerillos, Province of Mendoza, Argentina. A vouch- 370 M.L. Vaca Ruiz et al. er of each sample was deposited at the Herbario of the Uni- plates were allowed to dry for 5 min at room temperature. versidad Nacional de San Luis under numbers: 492- Del Sterilised paper disks (Britania) of 6 mm diameter were Vitto [A. echegarayi Hieron. (Asteraceae)], 8487-Del Vitto used. Ten microlitres of each essential oil and vegetable oil [F. oolepis Blake (Asteraceae)] and 9186-Del Vitto [B. sali- (negative control) were added to impregnate paper disks and cifolia (Ruiz & Pavon) Pers]. allowed dry for 15 min. Commercial gentamicin disks (10 µg, Britania) were used as positive control. The disks were then Extraction of essential oil. Fresh aerial parts (5000 g) of placed aseptically over the surface of the bacterial cultures each plant were cut into small pieces and subjected to on MH plates and incubated at 37 C for 24 h. After incuba- steam-distillation at 96 C for 3 h using a Clevenger type tion, the inhibition zones around paper disks were measured -1 apparatus. The oils obtained (0.56 g kg ) were dried over accurately using a metric ruler. Assays were carried out on anhydrous sodium sulphate (Deans and Svoboda, 1990). duplicate MH plates for each L. monocytogenes strain. The experiments were replicated at least twice and the data Cultures. Fourteen strains of L. monocytogenes, listed in were analysed using InfoStat program to calculate the vari- Table 1, were used for the study. This number includes three ance analysis. Significant differences in the antibacterial strains from the American Type Culture Collection and one effects between the essential oils studied were determined strain from the Pasteur Institute Collection. The remaining by Duncan test (P < 0.01). L. monocytogenes strains were isolated in our laboratory from human, animal, and food samples. Each strain of L. monocytogenes was individually cultured RESULTS AND DISCUSSION in Trypticase Soy broth (Britania, Argentina) at 37 C for 24 h. Following incubation, the bacterial cells were separated by The antibacterial effect of three essential oils on different L. centrifugation (5000 rpm for 15 min at 4 C), washed twice monocytogenes strains is shown in Table 1. The average inhi- with sterile phosphate buffered saline (PBS, pH 7.2), and bition zone of F. oolepis against L. monocytogenes ranged finally resuspended in PBS. The cultures were appropriate- from 18.7 ± 2.6 mm (L. monocytogenes CLIP 74903) to 8.7 ly diluted in PBS to obtain a concentration of 10 CFU/ml, (0.5 ± 0.9 mm (L. monocytogenes 4425), yielding a mean on Mac Farland scale). The bacterial population in each cul- inhibitory zone of 12.64 mm on L. monocytogenes strains. ture was verified by spreading 100 ml aliquots of the appro- The average inhibition zone of B. salicifolia ranged from priately diluted cultures on Trypticase Soy agar (Britania) 12.2 ±1.8 mm (L. monocytogenes CLIP 74903) to 8.0 ±0.7 plates, with incubation at 37 C for 24 h. mm (L. monocytogenes 4425), yielding a mean inhibitory zone of 10.63 mm on L. monocytogenes strains. Artemisia Determination of antibacterial effect. The antibacterial echegarayi essential oil showed the same inhibition pattern: effect of F. oolepis, B. salicifolia, and A. echegarayi on the 12.5 ± 1.1 mm (L. monocytogenes CLIP 74903) to 7.7 ± 1.0 fourteen strains of L. monocytogenes was determined by the mm (L. monocytogenes 4425), yielding a mean inhibitory standard disk diffusion technique (CLSI, 2005). A population zone of 9.56 mm on L. monocytogenes strains. Gentamicin of approximately 10 CFU/ml of each L. monocytogenes resulted in inhibition zones ranging from 33.5 ± 1.9 mm (L. strain was inoculated on duplicate plates containing Müeller monocytogenes I) to 23.8 ± 1.8 mm (L. monocytogenes Hinton agar (MH, Britania) using sterile cotton swabs. The 4425). Vegetable oil produced no L. monocytogenes inhibi- TABLE 1 – Inhibitory effect of Flourensia oolepis, Baccharis salicifolia, Artemisia echegarayi essential oils on Listeria monocytogenes strains a b c d e L. monocytogenes Source/origin GN (mm)* VO (mm) Fo (mm)* Bs (mm)** Ae (mm)** strains Mean ± SE Mean ± SE Mean ± SE Mean ± SE Mean ± SE A Human 30.0 ± 2.5 0 14.0 ± 2.9 10.4 ± 1.1 9.25 ± 0.6 D Animal 31.7 ± 2.0 0 11.5 ± 1.9 11.0 ± 1.4 8.50 ± 1.5 E Animal 30.0 ± 1.4 0 12.0 ± 1.8 10.5 ± 1.0 9.50 ± 0.6 G Food 29.4 ± 0.5 0 10.5 ± 2.3 9.00 ± 0.8 8.00 ± 1.0 H Food 29.2 ± 1.6 0 13.0 ± 1.4 10.5 ± 1.7 8.00 ± 1.1 I Food 33.5 ± 1.9 0 14.2 ± 3.7 12.0 ± 1.4 10.3 ± 0.7 K Food 30.2 ± 1.3 0 11.5 ± 1.0 10.2 ± 0.8 9.25 ± 0.6 MF2 Human 25.4 ± 2.3 0 9.70 ± 2.2 11.0 ± 1.0 10.0 ± 1.5 56 Food 29.2 ± 1.9 0 11.5 ± 2.3 9.40 ± 0.5 10.0 ± 0.6 4425 Human 23.8 ± 1.8 0 8.70 ± 0.9 8.00 ± 0.7 7.70 ± 1.0 35152 ATCC 30.2 ± 1.7 0 12.5 ± 0.7 11.3 ± 0.6 9.75 ± 0.0 74903 CLIP 31.8 ± 2.4 0 18.7 ± 2.6 12.2 ± 1.8 12.5 ± 1.1 19112 ATCC 30.2 ± 2.7 0 15.3 ± 0.6 11.2 ± 1.5 10.0 ± 2.1 19115 ATCC 29.5 ± 1.5 0 13.6 ± 0.6 12.0 ± 1.7 11.0 ± 2.8 a b c d e f GN: gentamicin, VO: vegetable oil, Fo: Fluorensia oolepis, Bs: Baccharis salicifolia, Ae: Artemisia echegarayi, ATCC: American Type Culture Collection, CLIP: Listeria Collection of the Pasteur Institute. * Very significative (P < 0.01), ** Significative (P < 0.05). Ann. Microbiol., 56 (4), 369-371 (2006) 371 tion zone, indicating that it does not have any antibacterial ty testing. Fifteenth Informational Supplement. Disk diffu- sion. M100-S15. Vol. 25 No. 1. effect on this pathogen. Although all strains of L. monocytogenes varied in their Cobos M., Rodriguez J., Oliva M., Demo M., Faillaci S., Zygadlo J. (2001). Composition and antimicrobial activity of the essen- antimicrobial susceptibility, F. oolepis exhibited the highest tial oil of Baccharis notosergila. Planta Med., 67: 84-86. activity with a highly significant difference as compared with Cosentino S., Tuberoso C., Pisano B., Satta M., Mascia V., Arzedi the other oils (P < 0.01). E., Palmas F. (1999). In vitro antimicrobial activity and chem- Gentamicin was used as a positive control in the study ical composition of Sardinian Thymus essential oils. Lett. Appl. because L. monocytogenes has been reported to be highly Microbiol., 29: 130-135. sensitive to this antibiotic. In fact, gentamicin and ampicillin Deans S., Svoboda K. (1990). The antimicrobial properties of are usually recommended for the treatment of listeriosis marjoram (Origanum majorana L.) volatile oil. Flavour Frag. (Hof et al., 1997). In a study on black seed (Nigella sativa) J., 5: 187-190. oil Nair et al. (2005) found that gentamicin inhibition zones Feresin G., Tapia A., Gimenez A., Gutierrez Ravelo A., Zacchino S., were smaller than those of black seed oil. The high activity Sortino M., Schmeda-Hirschman G. (2003). Constituents of the Argentinian medicinal plant Baccharis grisebachii and their of black seed oil is due to presence of carvacrol, thymol and antimicrobial activity. J. Ethnopharmacol., 89: 73-80. thymohydroquinone, which are well known for their anti- bacterial activities. 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Management of lis- methylethyl)-1,4-cyclohexadiene) is the precursor of thymol teriosis. Clin. Microbiol. Rev., 10: 345-357. (5-methyl-2-(1-methylethyl) phenol) (Cosentino et al., Juteau F., Massoti V., Bessiere J., Dherbomez M., Viano J. (2002). 1999). The essential oils evaluated in this work have a great Antibacterial and antioxidant activities of Artemisia annua variety of phytochemicals that could be considered respon- essential oil. Fitoterapia, 73: 532-535. sible for a larger or smaller part of the antibacterial activity. Kathariou S. (2002). Listeria monocytogenes virulence and patho- Although they usually occur as complex mixtures, their activ- genecity, a food safety perspective. J. Food Prot., 65: 1811- ity can be accounted for in terms of their major components. Research into the antimicrobial actions of monoterpenoids Laciar A.L., Centorbi O.N.P. de (2002). Listeria species in seafood: isolation and characterization of Listeria spp. from seafood in suggests that they diffuse into and damage cell membrane San Luis, Argentina. Food Microbiol., 19: 645-651. structures (Sikkema et al., 1995). Even though the exact Lundén J.M., Miettinen M.K., Autio T.J., Korkeala H.J. (2000). Per- nature of the antibacterial activity is not known, our results sistent Listeria monocytogenes strains show enhanced adher- indicate that the essential oils tested possess moderate ence to food contact surface after short contact times. J. Food activity against L. monocytogenes. However, appropriate Prot., 63: 1204-1207. applications in food should be validated. Nair M.K., Vasudevan P., Venkitanarayanan K. (2005). Antibacte- rial effect of black seed oil on Listeria monocytogenes. Food Acknowledgements Control, 16: 395-398. We have to thank to UNSL (Projects 7301 and 8802), CON- Setzer W., Vogler B., Schmidt J., Leahy J., Rives R. (2004). 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Annals of Microbiology – Springer Journals

Published: Nov 20, 2009

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