Action of Essential Oils Obtained from Baccharis coridifolia D. C. (Asteraceae-Astereae) on the Activity of Antibiotics

Sideney Becker Onofre, Marilde Canton, Paula Andressa Pires

.
Department of Biological Sciences and Pharmacy, FAED—Faculty of Educational of Dois Vizinhos—Uni?o de Ensino do Sudoeste do Paraná—UNISEP—Bairro Nossa Senhora Aparecida, Dois Vizinhos, Paraná, Brazil.

DOI: 10.4236/aim.2013.32026                

Abstract

The aim of this study was to investigate the effects of Baccharis coridifolia essential oil on the Activity of Antibiotics. Assays were performed with ampicillin (10 μg), cephalothin (30 μg), chloramphenicol (30 μg), gentamicin (10 μg) and tetracycline (30 μg) alone and in combination with the essential oil (4% v/v) through the disk diffusion susceptibility test. The results showed the effects of essential oil on the activity of the antibiotics tested. Zones of inhibition of bacterial growth with different diameters were observed surrounding the antibiotic disks, whether or not they were impregnated with the essential oil. The occurrence of the synergistic or antagonistic effect was observed in both bacterial strains assessed—Staphylococcus aureus (ATCC-25923) and Escherichia coli (ATCC-25922). These results show that the use of products derived from plants can, in some cases, interfere with the effectiveness of antibiotics during clinical therapy.

Keywords

Medicinal Plants; Essential Oils; Antibiotic; Associated Use; Interference

1. Introduction

The use of natural products as a therapeutic resource is as old as human civilization. Even with the growing economic power of the large pharmaceutical companies, natural products have not lost their place in therapy, being mistakenly regarded by the population as safe medicines and being increasingly used [1].

Plants with therapeutic properties used in traditional health care are an important source of new biologically active compounds. They have appeared as part of the traditional health care in many parts of the world for decades and have attracted the interest of several researchers [2-6].

In folk medicine, plants are used concomitantly with conventional medicaments [7]. In this combined use, medicinal plants and/or their byproducts may act by inhibiting or enhancing the therapeutic effect of conventional drugs, as well as not interacting in the expected way [8]. In dermatology, such associative use often puts the patient at risk, as it can trigger phytodermatitis due to irritation or photosensitivity mechanisms. This practice may also hinder the clinical diagnosis, as often the patients are not aware of the importance of informing health care professionals about the use of medicinal plants during a medical appointment.

Oils and plant extracts have long been used in numerous applications in folk medicine, including the production of topical antiseptics. This was the basis for several scientific investigations aimed at confirming the antimicrobial activity of essential oils [9-13].

Celiktas et al. [14] related the increase of antimicrobial activity of Rosmarinus officinalis essential oil in samples collected during the spring. Moon et al. [15] found significant variation in the size of inhibition zones when testing the antimicrobial activity of essential oils from different samples of Lavandula angustifolia against the same microorganisms.

The active ingredients present in plants have been extensively studied in many countries, because new drugs can be developed with this knowledge. Understanding the interactions between drugs and plants can also be useful in therapy (beneficial interaction) and for disease prevention.

Plants of the family Asteraceae are extensively studied for their chemical composition and biological activity, with some supporting the development of new drugs, insecticides and other products [16-18].

About 120 Baccharis species have been studied chemically with 30 of them investigated for biological activity. Generally, the most common compounds are the flavonoids and labdane and clerodane diterpenes, although kaurane diterpenes, triterpenes, germacrene, coumaric acids, trichothecenes, sesquiterpenes and phenylpropanoids are also frequently found. Allelopathic, antimicrobial, antiinflammatory and cytotoxic effects are highlighted in biological activity studies. The most researched species for chemical composition and/or biological activity include B. megapotamica, B. incarum, B. trimera, B. trinervis, B. salicifolia, B. crispa, B. coridifolia, B. dracunculifolia, B. uncinella, B. grisebachii and B. tricuneata [17- 20].

Assessing the biological activity of essential oils produced by B. dracunculifolia, B. uncinella and B. coridifolia Ferronato et al. [13] observed the antioxidant and anti-inflammatory effect of these oils on Escherichia coli ATCC, Staphylococcus aureus ATCC, Pseudomonas aeruginosa ATCC and Streptococcus mutans isolated from saliva, thus showing the importance of these plants. Besides the antimicrobial activity observed in several plant species, natural compounds can also exhibit synergism when used in conjunction with other drugs [21]. Thus, this study evaluated the effect of essential oil and extracts produced by B. coridifolia D. C. (Asteraceae) (Figure 1) on the activity of some antibiotics used in clinical therapy.

2. Materials and Methods

2.1. Botanical Material

The plant studied was B. coridifolia D. C. (Asteraceae) collected in the Southwest region of Paraná, Brazil, from

Figure 1. Baccharis coridifolia D. C.—Asteraceae-Astereae.

November 2006 to November 2007. The essential oils were obtained followed the method described by Onofre et al. [13,17,18,22]. This species was chosen because it is widely used in folk medicine and it contains antimicrobial substances described in previous studies [13,22].

2.2. Microorganisms

S. aureus (ATCC-25923) and E. coli (ATCC-25922) were used as test microorganisms. Suspensions were prepared in a 0.85% (w/v) NaCl solution, with turbidity level compared to the 0.5 McFarland standard, which approximately corresponds to a concentration of 10⁸ CFU/mL.

2.3. Antibiotics

Ampicillin (10 μg), cephalothin (10 μg), chloramphenicol (30 μg), gentamicin (10 μg) and tetracycline (30 μg) were selected in the study, because of their availability in primary healthcare. The observational study of bacterial susceptibility to antibiotics was carried out through the disk diffusion susceptibility test (NewprovÒ) [23].

2.4. Effect of Essential Oil on Antibiotic Activity

The disk diffusion susceptibility test was carried out to analyze the effect of the essential oil on antibiotic activity. The essential oil was studied at a 4% concentration, which corresponds to the MIC determined in previous studies for the microorganisms tested here [2-4,13,24-26]. The disks containing the antibiotics in their respective concentrations were impregnated with 20 μL of 4% essential oil (MIC), and then placed on sterile Petri plates containing Mueller-Hinton agar inoculated with 1 mL of bacterial suspensions. After incubating the plates at 37˚C for 48 h, the effect of the essential oil MIC on the activity of antibiotics was observed. Synergistic effect was considered when inhibition zones ≥2 mm diameter were observed surrounding disks containing the essential oil (EO) and the antibiotic (AB). Antagonistic effect was considered when an inhibition zone of smaller diameter was observed around the disks with AB and EO, compared to that developed by the AB disks. Indifferent effect was considered when there was a zone of inhibition surrounding the disks with AB and EO, with a diameter equal to that resulting from the application of the AB alone [27]. All assays were performed in duplicate and the results were obtained by averaging the results of the parallel tests.

3. Results and Discussion

Table 1 shows the antimicrobial susceptibility of the tested strains. The Gram-positive S. aureus was susceptible to all antibiotics tested, showing growth inhibition zones with diameters ranging from 12 to 24 mm. The Gram-negative E. coli was susceptible to cephalothin, gentamicin and tetracycline. These results are in agreement with the CSLI [28].

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	T. Eisenberg, S. Sundren and M. Wells, “Larger Board Size and Decreasing Firm Value in Small Firms,” Journal of Financial Economic, Vol. 48, No. 1, 1998, pp. 35-54. doi:10.1016/S0304-405X(98)00003-8
[2]	B. S. O. Ceballos, R. C. S. B. C. Barbosa, E. O. Lima and R. F. Urtiga, “Atividades Antimicrobianas de Produtos Naturais Sobre Staphylococcus aureus e Pseudomonas aeruginosa Isoladas de águas Recreacionais,” Brazilian Journal of Pharmacognosy, Vol. 74, No. 1, 1993, pp. 4-6.
[3]	E. O. Lima, “Estudo das Dermatofitoses em Joao Pessoa-Paraíba e da Atividade Antifungica de Plantas Medicinais da Regiso Contra Alguns de Seus Agentes Isolados,” Universidade de Sao Paulo, Sao Paulo, 1996.
[4]	E. O. Lima, N. M. P. Farias, E. L. Souza and B. H. C. Santos, “Propriedades Antibacterianas de óleos Essenciais de Plantas Medicinais,” Revista Brasileira de Ciencias da Saude, Vol. 7, No. 21, 2003, pp. 251-258.
[5]	G. M. A. Cunha, “Atividade Antimicrobiana de Plantas Popularmente Usadas no Ceará,” Brazilian Journal of Pharmacognosy, Vol. 76, No. 3, 1995, pp. 5-6.
[6]	M. N. Cowan, “Plant Products as Antimicrobial Agents,” Clinical Microbiology Reviews, Vol. 12, No. 4, 1999, pp. 564-582.
[7]	J. A. Amorim, “Fitoterapia Popular e Saude da Comunidade: Diagnostico Para Proposta de Integracao Nos Servicos de Saude em Campina Grande,” Universidade de Sao Paulo, Sao Paulo, Brazil, 1999.
[8]	G. F. Nascimento, J. Locatelli, P. C. Freitas and G. L. Silva, “Antibacterial Activity of Plant Extracts and Phytochemicals on Antibiotic-Resistant Bacteria,” Brazilian Journal of Microbiology, Vol. 31, No. 4, 2000, pp. 8-53.
[9]	D. H. Fine, D. Fergang, M. L. Barnett, C. Drew, L. Steinberg, C. H. Charles and J. W. Vincent, “Effect of an Essential Oil-Containing Antiseptic Mouthrinse on Plaque and Salivary Streptococcus mutans Levels,” Journal of Clinical Periodontology, Vol. 27, No. 3, 2000, pp. 157-161. doi:10.1034/j.1600-051x.2000.027003157.x
[10]	P. Pan, M. L. Barnett, J. Coelho, C. Brogdon and M. B. Finnegan, “Determination of the in Situ Bactericidal Activity of an Essential Oil Mouthrinse Using a Vital Stain Method,” Journal of Clinical Periodontology, Vol. 27, No. 4, 2000, pp. 256-261. doi:10.1034/j.1600-051x.2000.027004256.x
[11]	R. Seymour, “Additional Properties and Uses of Essential Oils,” Journal of Clinical Periodontology, Vol. 30, No. s5, 2003, pp. 19-21. doi:10.1034/j.1600-051X.30.s5.7.x
[12]	X. P. Nunes, G. L. A. Maia, J. R. G. S. Almeida, F. O. Pereira and E. O. Lima, “Antimicrobial Activity of the Essential Oil of Sida cordifolia L.,” Revista Brasileira de Farmacognosia, Vol. 16, No. 1, 2006, pp. 642-644. doi:10.1590/S0102-695X2006000500010
[13]	R. Ferronatto, E. D. Marchesan, F. Bednarski, S. M. Alencar and S. B. Onofre, “Atividade Antioxidante Dos óleos Essenciais, Produzidos por Baccharis dracunculifolia D.C. Baccharis uncinella e B. coridifolia D.C. (Asteraceae),” Arquivos de Ciências da Saúde da Unipar, Vol. 10, No. 2, 2007, pp. 67-70.
[14]	O. Y. Celiktas, E. E. H. Kocabas, E. Bedir, F. V. Sukan, T. Ozek and K. H. C. Baser, “Antimicrobial Activities of Methanol Extracts and Essential Oils of Rosmarinus officinalis, Depending on Location and Seasonal Variations,” Food Chemistry, Vol. 100, No. 2, 2006, pp. 553-559. doi:10.1016/j.foodchem.2005.10.011
[15]	T. Moon, J. M. Wilkinson and H. M. A. Cavanagh, “Antibacterial Activity of Essential Oils, Hidrosols an Plant Extracts from Australian Grown Lavandula spp.,” International Journal of Aromatherapy, Vol. 16, No. 1, 2006, pp. 9-14. doi:10.1016/j.ijat.2006.01.007
[16]	C. L. Queiroga, A. E. Fukai and A. Marsaioli, “Composition of the Essential Oil of Vassoura,” Journal of Brazilian Chemical Society, Vol. 1, No. 4, 1990, pp. 105-109. doi:10.5935/0103-5053.19900023
[17]	S. B. Onofre, R. Ferronatto, E. D. Marchesan and F. Bednarski, “Avaliacao da Atividade Antimicrobiana de oleos Essenciais Produzidos por B. dracunculifolia e B. uncinella, Obtidos da Regiao Sudoeste do Paraná,” Anais, 2006, pp. 46-52.
[18]	S. B. Onofre, R. Ferronatto, E. D. Marchesan and F. Bednarski, “Avaliacao da Atividade Antioxidante de oleos Essenciais Produzidos por B. dracunculifolia e B. uncinella, Obtidos da Regiao Sudoeste do Paraná,” Anais, 2006, pp. 72-78.
[19]	I. Loayza, G. Collin, M. Gagnon, H. Deslauriers and E. Dellacassa, “Huiles Essentielles de Baccharis latifolia, B. salicifolia de Bolivia et de B. dracunculifolia en Provenance d’Uruguay,” Rivista Italiana, Vol. 12, 1995, pp. 28-35.
[20]	L. G. Verdi, I. M. C. Brighente and M. G. Pizzolatti, “O Gênero Baccharis (Asteraceae): Aspectos Químicos, Economicos e Biologicos,” Quimica Nova, Vol. 28, No. 1, 2005, pp. 85-94. doi:10.1590/S0100-40422005000100017
[21]	J. C. Sidrim and J. L. B. Moreira, “Fundamentos Clínicos e Laboratoriais da Micologia Medica,” Guanabara Koogan, Rio de Janeiro, 1999.
[22]	E. D. Marchesan, R. Ferronatto, F. Bednarski, S. M. Alencar and S. B. Onofre, “Acao dos óleos Essenciais Produzidos por Baccharis dracunculifolia D.C. e Baccharis uncinella D.C. (Asteraceae) Sobre a Atividade Hialuronidase,” Arquivos de Ciências da Saude da Unipar, Vol. 10, No. 2, 2006, pp. 63-66.
[23]	A. W. Bauer, M. M. Kirby, J. C. Sherris and M. Truck, “Antibiotic Susceptibility Testing by a Standardized Single Disk Method,” American Journal of Clinical Pathology, Vol. 45, No. 4, 1966, pp. 493-496.
[24]	J. Allegrini, M. S. Bouchberg and H. Maillols, “Emulsions d’huiles Essentielles Fabrication et Applications em Microbiologie,” Societe de Pharmacie de Montpellier, Vol. 33, No. 1,1973, pp. 73-86.
[25]	J. C. L. V. Araujo, E. O. Lima, B. S. Ceballos, K. R. L. Freire, E. L. Souza and L. Santos-Filho, “Acao Antimicrobiana de óleos Essenciais Sobre Microrganismos Potencialmente Causadores de Infeccoes Oportunistas,” Revista de Patologia Tropical, Vol. 33, No. 1, 2004, pp. 55-64.
[26]	J. C. R. Nogueira, “Atividade Antimicrobiana in Vitro de Produtos Vegetais Sobre Microrganismos Isolados de Pacientes Portadores de Otite Externa Aguda,” Universidade Federal da Paraíba, Paraíba, 2004.
[27]	L. Cleeland and E. Squires, “Evaluation of New Antimicrobials in Vitro and Experimental Animal Infections,” In: V. M. D. Lorian, Ed., Antibiotics in Laboratory Medicine, Williams & Wilkins, Baltimore, 1991, pp. 739-788.
[28]	CLSI—National Committee for Clinical Laboratory Standards—Approved standard M7-A5, “Methods for Dilution Antimicrobial Susceptibility Test for Bacteria That Grow Aerobicaly,” 5th Edition, CLSI, 2003.
[29]	M. Canton and S. B. Onofre, “Interferência de Extratos da Baccharis dracunculifolia D.C., Asteraceae, Sobre a Atividade de Antibióticos Usados na Clínica,” Brazilian Journal of Pharmacognosy, Vol. 20, No. 3, 2009, pp. 348-354.
[30]	D. Araujo and S. B. Onofre, “Acao do Extrato Hidroalcoólico de Alternanthera brasiliana (L.) O. Kunt., (Amaranthaceae) Sobre a Atividade de Antimicrobianos Utilizados na Terapeutica,” Sabios, Vol. 6, No. 1, 2011, pp. 23-26.