Evaluation of the antibacterial effect of (R)-(+)-Limonene against Enterococcus faecalis and Enterobacter cloacae strains isolated from food

Authors

DOI:

https://doi.org/10.5433/1679-0359.2024v45n4p1201

Keywords:

Antibacterial activity, Antimicrobials, Phytotherapy, Monoterpene, Natural Products.

Abstract

The aim of this study was to evaluate the potential antibacterial and anti-adherent activities of the monoterpene (R)-(+)-limonene, as well as its synergistic potential with synthetic antimicrobials against strains of Enterococcus faecalis and Enterobacter cloacae. The antibacterial properties of (R)-(+)-limonene were assessed using the broth microdilution technique to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Additionally, the infusion disc method was employed to explore the association of the compound with antimicrobials, and the test tube method was used to determine the minimum inhibitory concentration of adherence (MICA). It was observed that the MIC for (R)-(+)-limonene was 1000 μg mL-1 for five of the six E. faecalis strains tested, while for E. cloacae, the MIC exceeded 1000 μg mL-1 for all strains tested. Identical values were recorded for the MBC in E. faecalis. In terms of its combination with synthetic antimicrobials, (R)-(+)-limonene demonstrated a synergistic effect with gentamicin and ciprofloxacin for most strains. Regarding the MICA, both (R)-(+)-limonene and 0.12% chlorhexidine digluconate failed to inhibit biofilm formation at the tested concentrations. Given the need for new therapeutic alternatives for treating bacterial infections, this study revealed that the tested monoterpene exhibited moderate bactericidal effects against E. faecalis strains and no antibacterial effect against E. cloacae strains. However, when combined with various classes of antimicrobials, (R)-(+)-limonene showed synergistic effects with gentamicin and ciprofloxacin for most strains. This suggests that (R)-(+)-limonene holds promise for enhancing the treatment of bacterial infections and could support conventional therapies. Nonetheless, further in vitro, ex vivo, and in vivo studies are necessary to confirm and elucidate its efficacy and mechanisms.

 

Author Biographies

Millena de Souza Alves, Universidade Federal de Campina Grande

Doctoral Student in the Postgraduate Program in Animal Science and Health, Universidade Federal de Campina Grande, UFCG, Patos, PB, Brazil.

Maria Alice Araújo de Medeiros , Universidade Federal de Campina Grande

Doctoral Student in the Postgraduate Program in Animal Science and Health, Universidade Federal de Campina Grande, UFCG, Patos, PB, Brazil.

Bernadete Santos, Universidade Federal de Campina Grande

Doctoral Student in the Postgraduate Program in Animal Science and Health, Universidade Federal de Campina Grande, UFCG, Patos, PB, Brazil.

Mylena Medeiros Simões, Universidade Federal de Campina Grande

Master's Student in the Postgraduate Program in Animal Science and Health, UFCG, Patos, PB, Brazil.

João Henrique Anizio de Farias, Universidade Federal de Campina Grande

Master's Student in the Postgraduate Program in Animal Science and Health, UFCG, Patos, PB, Brazil.

Hilzeth de Luna Freire Pessôa, Universidade Federal da Paraíba

Profa. Dra.,, Universidade Federal da Paraíba, UFPB, João Pessoa, PB, Brazil.

Veneziano Guedes de Sousa Rêgo , Universidade Federal de Campina Grande

Prof. Dr., UFCG, Patos, PB, Brazil.

Raline Mendonça dos Anjos , Universidade Federal de Campina Grande

Profa. Dra., UFCG, Patos, PB, Brazil.

Luciano de Brito Junior , Universidade Federal de Campina Grande

Prof. Dr., UFCG, Patos, PB, Brazil.

Abrahão Alves de Oliveira Filho, Universidade Federal de Campina Grande

Prof. Dr. in the Postgraduate Program in Animal Science and Health, UFCG, Patos, PB, Brazil.

References

Albuquerque, A. C. L., Pereira, M. D. S. V., Pereira, J. V., Pereira, L. F., Silva, D. F., Macedo-Costa, M. R., & Higino, J. S. (2010). Anti-adherent effect of Matricaria extract recut Linn. on dental biofilm microorganisms. Revista de Odontologia da UNESP, 39(1), 21-25.

Almeida, S., Jr., Pereira, D. V., Ferreira, T. M., Freitas, R. A., Silva, C. C., Santos, M. F. C., Borges, C. H. G., Silva, M. A., Ambrósio, A. R., Bastos, J. K., Ross, S. A., & Furtado, Y. R. A. (2020). Anti-inflammatory and antinociceptive effects of kaempferide from the Brazilian green propolis. Research, Society and Development, 9(10), 1259108232. doi: 10.33448/rsd-v9i10.8232

Badke, M. R., Budó, M. D. L. D., Alvim, N. A. T., Zanetti, G. D., & Heisler, E. V. (2012). Knowledge and popular health care practices with the use of medicinal plants. Texto & Contexto-Enfermagem, 21(2), 363-370. doi: 10.1590/S0104-07072012000200014

Bauer, A. W., Kirby, W. M., Sherris, J. C., & Turck, M. (1996). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45(4), 493-496. doi: 10.1093/ajcp/45.4_ts.493

Bennett, J. E., Dolin, R., & Blaser, M. J. (2019). Mandell, Douglas, and Bennett's principles and practice of infectious diseases E-Book: 2-volume set. Elsevier Health Sciences.

Berthe, F. C. J., Bouley, T., Karesh, W. B., Le Gall, F. G., Machalaba, C. C., Plante, C. A., & Seifman, R. M. (2018). Operational framework for strengthening human, animal and environmental public health systems at their interface. World Bank Group.

Bona, E. A. M. D., Pinto, F. G. D. S., Fruet, T. K., Jorge, T. C. M., & Moura, A. C. D. (2014). Comparison of methods for the evaluation of antimicrobial activity and determination of the minimum inhibitory concentration (mim) of aqueous and ethanolic plant extracts. Arquivos do Instituto Biológico, 81(3), 218-225. doi: 10.1590/1808-1657001192012

Braga, L. M. P. S., Souza, V. T., Rodrigues, M. M., Ramos, G. L. D. P. A., & Gomes, A. D. S. G. (2020). Analyze juice microbiological green commercialized per one network retailer at city of Rio de Janeiro. Food Science and Technology, 1, 29-47.

Chajęcka-Wierzchowska, W., Zadernowska, A., & Laniewska-Trokenheim, L. (2017). Virulence factors of Enterococcus spp. presented in food. LWT, 75, 670-676. doi: 10.1016/j.lwt.2016.10.026

Chikhoune, A., Hazzit, M., Kerbouche, L., Baaliouamer, A., & Aissat, K. (2013). Tetraclinis articulata (Vahl) Masters essential oils: chemical composition and biological activities. Journal of Essential Oil Research, 25(4), 300-307. doi: 10.1080/10412905.2013.774625

Cleeland, R., & Squires, E. (1991). Evaluation of new antimicrobials in vitro and in experimental animal infections. Antibiotics in Laboratory Medicine, 3, 739-787.

Clinical and Laboratory Standards Institute (2012). Dilution antimicrobial susceptibility testing methods for aerobically growing bacteria (9nd ed.). (Document CLSI MO7- A9). CLSI.

Costa, M. D. S., Rocha, J. E., Campina, F. F., Silva, A. R., Cruz, R. P., Pereira, R. L., & Coutinho, H. D. (2019). Comparative analysis of the antibacterial and drug-modulatory effect of d-limonene alone and complexed with β -cyclodextrin. European Journal of Pharmaceutical Sciences, 128, 158-161. doi: 10.1016/j.ejps.2018.11.036

Davin-Regli, A., Lavigne, J. P., & Pagès, J. M. (2019). Enterobacter spp.: update on taxonomy, clinical aspects, and emerging antimicrobial resistance. Clinical Microbiology Reviews, 32(4), 10-1128. doi: 10.1128/cmr.00002-19

Farmer, T. J., Comerford, J. W., Pellis, A., & Robert, T. (2018). Post‐polymerization modification of bio‐based polymers: maximizing the high functionality of polymers derived from biomass. Polymer International, 67(7), 775-789. doi: 10.1002/pi.5573

Flores, A. M. P. C., & Melo, C. B. (2015). Main bacteria that cause foodborne illness. Revista Brasileira de Medicina Veterinária, 37(1), 65-72.

Guerra, F. Q. S., Mendes, J. M., Oliveira, W., Costa, J., Coutinho, H. D. M., & Lima, E. O. (2012). Chemical composition and antimicrobial activity of Cinnamomum zeylanicum Blume essential oil on multi-drug resistant Acinetobacter spp. Strains. Revista de Biologia e Farmacia, 8(1), 62-70.

Hafidh, R. R., Abdulamir, A. S., Vern, L. S., Abu Bakar, F., Abas, F., Jahanshiri, F., & Sekawi, Z. (2011). Inhibition of growth of highly resistant bacterial and fungal pathogens by a natural product. The Open Microbiology Journal, 5(1), 96-106. doi: 10.2174/1874285801105010096

Langeveld, W. T., Veldhuizen, E. J., & Burt, S. A. (2014). Synergy between essential oil components and antibiotics: a review. Critical Reviews in Microbiology, 40(1), 76-94. doi: 10.3109/1040841X.2013.763219

Lebreton, F., Willems, R. J. L., & Gilmore, M. S. (2014). Diversity of Enterococcus, origins in nature and intestinal colonization. In M. Gilmore, D. Clewell, Y. Ike, & N. Shankar (Eds.), Enterococci; from commensals to major causes of drug resistant infection (pp. 1-59). Boston.

Milenković, M., Stošović, J., & Slavkovska, V. (2018). Synergy between essential oils of Calamintha species (Lamiaceae) and antibiotics. Natural Product Communications, 13(3), 1934578X1801300325. doi: 10.1177/1934578X1801300325

Nacu, A., & Hoerr, R. (2016). Neuropsychiatric symptoms in dementia and the effects of Ginkgo biloba extract EGb 761® treatment: additional results from a 24-week randomized, placebo-controlled trial. Open Access Journal of Clinical Trials, 8, 1-6. doi: 10.2147/OAJCT.S93531

Ncube, N., Afolayan, S. A. J., & Okoh, A. I. (2008). Assessment techniques of antimicrobial properties of natural compounds of plant origin: Current methods and future trends. African Journal of Biotechnology, 7(12), 1797-1806. doi: 10.5897/AJB07.613

Oliveira, R. A. G., Lima, E. O., Vieira, W. L., Freire, K. R. L., Trajano, V. N., Lima, I. O., Souza, E. L., Toledo, M. S., & Silva, R. N., Fº. (2006). Estudo da interferência de óleos essenciais sobre a atividade de alguns antibióticos usados na clínica. Revista Brasileira de Farmacognosia, 16(1), 77-82. doi: 10.1590/S0102-695X2006000100014

Ostrosky, E. A., Mizumoto, M. K., Lima, M. E. L., Kaneko, T. M., Nishikawa, S. O., & Freitas, B. R. (2008). Métodos para avaliação da atividade antimicrobiana de determinação de concentração mínima inibitória (CMI) de plantas medicinais. Revista Brasileira de Farmacognosia, 18(2), 301-307. doi: 10.1590/S0102-695X2008000200026

Palomino, J. C., Martin, A., Camacho, M., Guerra, H., Swings, J., & Portales, F. (2002). Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistence Mycobacterium tuberculosis. Antimicrobial Agents and Chemotherapy, 46(8), 2720-2722. doi: 10.1128/aac.46.8.2720-2722.2002

Retajczyk, M., & Wróblewska, A. (2019). Isomerization and dehydroaromatization of R (+)-limonene over the Ti-MCM-41 catalyst: effect of temperature, reaction time and catalyst content on product yield. Catalysts, 9(6), 508. doi: 10.3390/catal9060508

Sánchez Cabrera, A., Parada, R., Marguet, E., & Vallejo, M. (2021). Virulence factors, antimicrobial resistance ya heavy metals in Enterococcus spp. isolated from foods of animal origin. Acta Bioquímica Clínica Latinoamericana, 55(2), 127-135.

Santana, M. T. P., Santos, T. A., Gomes, L. L., Alves, M. A. S. G., Nogueira, P. L., Dantas, M. V. O., Silva, Q. P., Medeiros, L. A. D. M., Rosendo, R. A., Anjos, R. M., Sousa, M. A. N., Brito, L., Jr., Guênes, G. M. T., Penha, E. S., Almeida, M. S. C., Sousa, A. P., & Oliveira, A. A., Fº. (2021). Antibacterial activity of Lavandula essential oil hybrida Grosso associated with cephalothin against strains of Staphylococcus aureus. Research, Society and Development, 10(2), e47110212682-e47110212682. doi: 10.33448/rsd-v10i2.12682

Sartoratto, A., Machado, A. L. M., Delarmelina, C., Figueira, G. M., Duarte, M. C. T., & Rehder, V. L. G. (2004). Composition and antimicrobial activity of essential oils from aromatic plants used in Brazil. Brazilian Journal of Microbiology, 35(4), 275-280. doi: 10.1590/S1517-83822004000300001

Settanni, L., Palazzolo, E., Guarrasi, V., Aleo, A., Mammina, C., Moschetti, G., & Germanà, M. A. (2012). Inhibition of foodborne pathogen bacteria by essential oils extracted from citrus fruits cultivated in Sicily. Food Control, 26(2), 326-330. doi: 10.1016/j.foodcont.2012.01.050

Silva, E. G., Bandeira, G., Jr., Cargnelutti, J. F., Santos, R. C. V., Gündel, A., & Baldisserotto, B. (2021). In vitro antimicrobial and antibiofilm activity of S-(-)-limonene and R-(+)-limonene against fish bacteria. Fishes, 6(3), 32. doi: 10.3390/fishes6030032

Silva, M. O., & Aquino, S. (2018). Resistência aos antimicrobianos: uma revisão dos desafios na busca por novas alternativas de tratamento. Revista de Epidemiologia e Controle de Infecção, 8(4), 472-482. doi: 10.17058/reci.v8i4.11580

Siqueira, I. N., Cavalcante, A. A. C., Sousa, D. L. C., Aquino, V. V. F., Oliveiro, L. B. S. de, Silva, J. G., Santos, C. S. A. B., Mota R. A., Oliveira A. A. Fo., & Melo, M. A. (2022). Isolamento e perfil de resistência antimicrobiana de Enterococcus spp em linhas de processamento de leite de cabra. Research, Society and Development, 11(9), e8611931269. doi: 10.33448/rsd-v11i9.31269

Souza, E. R. L., Araújo, C. J. H., Ferreira, J. L. S., Oliveira, H. M. B. F., & Oliveira, A. A., Fº. (2021). Antimicrobial and anti-adherent potential of Lavandula hybrida grosso essential oil against Klebsiella pneumoniae strains. Archives of Health Investigation, 10(6), 906-912. doi: 10.21270/archi.v10i6.5008

Souza, S. P., Cardoso, M. G., Souza, P. E., Guimarães, L. G. L., Andrade, J., Mallet, A. C. T., & Nelson, D. L. (2011). Baccharis essential oil tridentata Vahl; chemical composition, antioxidant and fungitoxic activity, and morphological characterization of secretory structures by scanning electron microscopy. Revista Brasileira de Plantas Medicinais, 13(4), 456-466. doi: 10.1590/S1516-05722011000400011

Sun, J. (2007). D-limonene: safety and clinical applications. Alternative Medicine Review, 12(3), 259.

Weiner, L. M., Webb, A. K., Limbago, B., Dudeck, M. A., Patel, J., Kallen, A. J., Edwards, J. R., & Sievert, D. M. (2016). Antimicrobial-resistant pathogens associated with healthcare-associated infections: summary of data reported to the national healthcare safety network at the centers for disease control and prevention, 2011–2014. Infection Control & Hospital Epidemiology, 37(11), 1288-1301. doi: 10.1017/ice.2016.174

Werner, G., Coque, T. M., Franz, C. M., Grohmann, E., Hegstad, K., Jensen, L., Schaik, W., & Weaver, K. (2013). Antibiotic resistant enterococci - tales of a drug resistance gene trafficker. International Journal of Medical Microbiology, 303(6-7), 360-379. doi: 10.1016/j.ijmm.2013.03.001

Downloads

Published

2024-07-31

How to Cite

Alves, M. de S., Medeiros , M. A. A. de, Santos, B., Simões, M. M., Farias, J. H. A. de, Pessôa, H. de L. F., … Oliveira Filho, A. A. de. (2024). Evaluation of the antibacterial effect of (R)-(+)-Limonene against Enterococcus faecalis and Enterobacter cloacae strains isolated from food. Semina: Ciências Agrárias, 45(4), 1201–1214. https://doi.org/10.5433/1679-0359.2024v45n4p1201

Issue

Section

Articles

Most read articles by the same author(s)

Similar Articles

You may also start an advanced similarity search for this article.