Investigating the antimicrobial activity of neem and clove extract on biofilm-producing oral microflora

Authors

DOI:

https://doi.org/10.18006/2024.12(3).511.521

Keywords:

Antifungal effect, Clove, Neem, Biofilm, Oral microbiota, Periodontal diseases

Abstract

Periodontal disease, a serious gum infection, is reported to be widespread in the Indian population. A heterogeneous microbial population, predominantly consisting of gram-negative anaerobes such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Porphyromonas intermedia, is associated with periodontal plaque formation. This condition may be worsened by the invasion of soft tissues by different species of Candida. Natural products like clove (Syzygium aromaticum) and Neem (Azadirachta indica) are very popular and easily available in the Indian climate and have great potential in preventing periodontitis. Azadirachta indica (Neem) exhibits versatile modes of action, including reported antimicrobial effects against several species associated with periodontal disease. Therefore, this study aims to detect the antimicrobial and antifungal effects of Neem and clove on oral biofilm both before and after biofilm formation. Results of the study revealed that both neem and clove crude extracts and their different dilution showed a significant reduction in the growth of fungal strains (Candida sp.) isolated from oral samples from people with poor hygiene and the biofilm produced by them.

References

Aas, J. A., Paster, B. J., Stokes, L. N., Olsen, I., & Dewhirst, F. E. (2005). Defining the normal bacterial flora of the oral cavity. Journal of Clinical Microbiology, 43(11), 5721–5732. https://doi.org/10.1128/JCM.43.11.5721-5732.2005 DOI: https://doi.org/10.1128/JCM.43.11.5721-5732.2005

Afolabi, O. C., Ogunsola, F. T., & Coker, A. O. (2008). Susceptibility of cariogenic Streptococcus mutans to extracts of Garcinia kola, Hibiscus sabdariffa, and Solanum americanum. West African journal of medicine, 27(4), 230–233.

Agarwal, V., Lal, P., & Pruthi, V. (2010). Effect of plant oils on Candida albicans. Journal of Microbiology, Immunology and Infection, 43(5), 447-451. DOI: https://doi.org/10.1016/S1684-1182(10)60069-2

Al-Ahmad, A., Ameen, H., Pelz, K., Karygianni, L., Wittmer, A., et al. (2014). Antibiotic resistance and capacity for biofilm formation of different bacteria isolated from endodontic infections associated with root-filled teeth. Journal of Endodontics, 40(2), 223-230. DOI: https://doi.org/10.1016/j.joen.2013.07.023

Almas K. (1999). The antimicrobial effects of extracts of Azadirachta indica (Neem) and Salvadora persica (Arak) chewing sticks. Indian journal of dental research: official publication of Indian Society for Dental Research, 10(1), 23–26.

Amanpour, S., AkbariJavar, M., Sarhadinejad, Z., Doustmohammadi, M., Moghadari, M., & Sarhadynejad, Z. (2023). A systematic review of medicinal plants and herbal products' effectiveness in oral health and dental cure with health promotion approach. Journal of Education and Health Promotion, 12, 306. https://doi.org/10.4103/jehp.jehp_1297_22 DOI: https://doi.org/10.4103/jehp.jehp_1297_22

Antimicrobial Resistance Collaborators (2024). The burden of bacterial antimicrobial resistance in the WHO African region in 2019: a cross-country systematic analysis. The Lancet Global Health, 12(2), e201–e216. https://doi.org/10.1016/S2214-109X(23)00539-9 DOI: https://doi.org/10.1016/S2214-109X(23)00539-9

Arweiler, N. B., & Netuschil, L. (2016). The Oral Microbiota. Advances in experimental medicine and biology, 902, 45–60. https://doi.org/10.1007/978-3-319-31248-4_4 DOI: https://doi.org/10.1007/978-3-319-31248-4_4

Bansal, V., Gupta, M., Bhaduri, T., Shaikh, S. A., Sayed, F. R., Bansal, V., & Agrawal, A. (2019). Assessment of antimicrobial effectiveness of neem and clove extract against Streptococcus mutans and Candida albicans: an in vitro study. Nigerian Medical Journal: Journal of the Nigeria Medical Association, 60(6), 285. DOI: https://doi.org/10.4103/nmj.NMJ_20_19

Barua, D. R., Basavanna, J. M., & Varghese, R. K. (2017). Efficacy of neem extract and three antimicrobial agents incorporated into tissue conditioner in inhibiting the growth of C. albicans and S. mutans. Journal of Clinical and Diagnostic Research, 11(5), ZC97. DOI: https://doi.org/10.7860/JCDR/2017/23784.9950

Batiha, G. E., Alkazmi, L. M., Wasef, L. G., Beshbishy, A. M., Nadwa, E. H., & Rashwan, E. K. (2020). Syzygium aromaticum L. (Myrtaceae): Traditional Uses, Bioactive Chemical Constituents, Pharmacological and Toxicological Activities. Biomolecules, 10(2), 202. https://doi.org/10.3390/biom10020202 DOI: https://doi.org/10.3390/biom10020202

Ben Hassine, D., Kammoun El Euch, S., Rahmani, R., Ghazouani, N., Kane, R., Abderrabba, M., &Bouajila, J. (2021).Clove Buds Essential Oil: The Impact of Grinding on the Chemical Composition and Its Biological Activities Involved in Consumer's Health Security. BioMed research international, 2021, 9940591. https://doi.org/10.1155/2021/9940591 DOI: https://doi.org/10.1155/2021/9940591

Buakaew, W., Sranujit, R. P., Noysang, C., Sangouam, S., Suphrom, N., Thongsri, Y., Potup, P., & Usuwanthim, K. (2021). Evaluation of Mouthwash Containing Citrus hystrix DC., Moringa oleifera Lam. and Azadirachta indica A. Juss. Leaf Extracts on Dental Plaque and Gingivitis. Plants (Basel, Switzerland), 10(6), 1153. https://doi.org/10.3390/plants10061153 DOI: https://doi.org/10.3390/plants10061153

Caselli, E., Fabbri, C., D'Accolti, M., Soffritti, I., Bassi, C., Mazzacane, S., & Franchi, M. (2020). Defining the oral microbiome by whole-genome sequencing and resistome analysis: the complexity of the healthy picture. BMC Microbiology, 20(1), 120. https://doi.org/10.1186/s12866-020-01801-y DOI: https://doi.org/10.1186/s12866-020-01801-y

Cruz Martínez, C., Diaz Gómez, M., & Oh, M. S. (2017). Use of traditional herbal medicine as an alternative in dental treatment in Mexican dentistry: a review. Pharmaceutical Biology, 55(1), 1992-1998. DOI: https://doi.org/10.1080/13880209.2017.1347188

Cugini, C., Ramasubbu, N., Tsiagbe, V. K., & Fine, D. H. (2021).Dysbiosis from a microbial and host perspective relative to oral health and disease. Frontiers in microbiology, 12, 617485. DOI: https://doi.org/10.3389/fmicb.2021.617485

D'Ambrosio, F., Santella, B., Di Palo, M. P., Giordano, F., & Lo Giudice, R. (2023). Characterization of the Oral Microbiome in Wearers of Fixed and Removable Implant or Non-Implant-Supported Prostheses in Healthy and Pathological Oral Conditions: A Narrative Review. Microorganisms, 11(4), 1041. https://doi.org/10.3390/microorganisms11041041 DOI: https://doi.org/10.3390/microorganisms11041041

Danthu, P., Simanjuntak, R., Fawbush, F., Tsy, J. L. P., Razafimamonjison, G., et al. (2020). The clove tree and its products (clove bud, clove oil, eugenol): prosperous today but what of tomorrow's restrictions?. Fruits, 75(5), 224-242. DOI: https://doi.org/10.17660/th2020/75.5.5

Deo, P. N., & Deshmukh, R. (2019). Oral microbiome: Unveiling the fundamentals. Journal of oral and maxillofacial pathology: JOMFP, 23(1), 122–128. https://doi.org/10.4103/jomfp.JOMFP_ 304_18 DOI: https://doi.org/10.4103/jomfp.JOMFP_304_18

Dorman, H. J., & Deans, S. G. (2000). Antimicrobial agents from plants: antibacterial activity of plant volatile oils. Journal of Applied Microbiology, 88(2), 308–316. https://doi.org/10.1046/ j.1365-2672.2000.00969.x DOI: https://doi.org/10.1046/j.1365-2672.2000.00969.x

Gasner, N. S., & Schure, R. S. (2023).Periodontal Disease. In Stat Pearls. StatPearls Publishing.

Ghannoum, M. A., Jurevic, R. J., Mukherjee, P. K., Cui, F., Sikaroodi, M., Naqvi, A., & Gillevet, P. M. (2010). Characterization of the oral fungal microbiome (mycobiome) in healthy individuals. PLoS pathogens, 6(1), e1000713. https://doi.org/10.1371/journal.ppat.1000713 DOI: https://doi.org/10.1371/journal.ppat.1000713

Hajishengallis, G., & Chavakis, T. (2021).Local and systemic mechanisms linking periodontal disease and inflammatory comorbidities. Nature Reviews. Immunology, 21(7), 426–440. https://doi.org/10.1038/s41577-020-00488-6 DOI: https://doi.org/10.1038/s41577-020-00488-6

Harjai, K., Bala, A., Gupta, R. K., & Sharma, R. (2013). Leaf extract of Azadirachta indica (Neem): a potential antibiofilm agent for Pseudomonas aeruginosa. Pathogens and Disease, 69(1), 62-65. DOI: https://doi.org/10.1111/2049-632X.12050

Janakiram, C., Mehta, A., & Venkitachalam, R. (2020). Prevalence of periodontal disease among adults in India: A systematic review and meta-analysis. Journal of Oral Biology and Craniofacial Research, 10(4), 800-806. DOI: https://doi.org/10.1016/j.jobcr.2020.10.016

John, J. R., Daniel, B., Paneerselvam, D., & Rajendran, G. (2017). Prevalence of Dental Caries, Oral Hygiene Knowledge, Status, and Practices among Visually Impaired Individuals in Chennai, Tamil Nadu. International journal of dentistry, 2017, 9419648. https://doi.org/10.1155/2017/9419648 DOI: https://doi.org/10.1155/2017/9419648

Kitamoto, S., Nagao-Kitamoto, H., Hein, R., Schmidt, T. M., & Kamada, N. (2020).The Bacterial Connection between the Oral Cavity and the Gut Diseases. Journal of dental research, 99(9), 1021–1029. https://doi.org/10.1177/0022034520924633 DOI: https://doi.org/10.1177/0022034520924633

Könönen, E., Gursoy, M., & Gursoy, U. K. (2019). Periodontitis: A Multifaceted Disease of Tooth-Supporting Tissues. Journal of Clinical Medicine, 8(8), 1135. https://doi.org/10.3390/jcm8081135 DOI: https://doi.org/10.3390/jcm8081135

Lamont, R. J., Koo, H., & Hajishengallis, G. (2018). The oral microbiota: dynamic communities and host interactions. Nature Reviews Microbiology, 16(12), 745-759. DOI: https://doi.org/10.1038/s41579-018-0089-x

Li X, Liu Y, Yang X, Li C, Song Z. (2022) The Oral Microbiota: Community Composition, Influencing Factors, Pathogenesis, and Interventions. Frontiers in Microbiology, 13, 895537.doi: 10.3389/fmicb.2022.895537. DOI: https://doi.org/10.3389/fmicb.2022.895537

Liu, K. C. S. C., Yang, S. L., Roberts, M. F., Elford, B. C., & Phillipson, J. D. (1992). Antimalarial activity of Artemisia annua flavonoids from whole plants and cell cultures. Plant Cell Reports, 11, 637-640. DOI: https://doi.org/10.1007/BF00236389

Maggini, V., Semenzato, G., Gallo, E., Nunziata, A., Fani, R., & Firenzuoli, F. (2024). Antimicrobial Activity of Syzygium aromaticum Essential Oil in Human Health Treatment. Molecules (Basel, Switzerland), 29(5), 999. https://doi.org/10.3390/ molecules29050999 DOI: https://doi.org/10.3390/molecules29050999

Marchese, A., Barbieri, R., Coppo, E., Orhan, I. E., Daglia, M., et al. (2017). Antimicrobial activity of eugenol and essential oils containing eugenol: A mechanistic viewpoint. Critical Reviews in Microbiology, 43(6), 668–689. https://doi.org/10.1080/ 1040841X.2017.1295225 DOI: https://doi.org/10.1080/1040841X.2017.1295225

Mathur, A., Dua, V.K., & Prasad, GBKS (2010) Antimicrobial Activity of Leaf Extracts of Murraya koenigii against Aerobic Bacteria Associated with Bovine Mastitis. International Journal of Chemical, Environmental and Pharmaceutical Research, 1(1), 12– 16.

Milutinovici, R. A., Chioran, D., Buzatu, R., Macasoi, I., Razvan, S., et al. (2021). Vegetal Compounds as Sources of Prophylactic and Therapeutic Agents in Dentistry. Plants (Basel, Switzerland), 10(10), 2148. https://doi.org/10.3390/plants10102148 DOI: https://doi.org/10.3390/plants10102148

Mosaddad, S. A., Hussain, A., & Tebyaniyan, H. (2023). Green Alternatives as Antimicrobial Agents in Mitigating Periodontal Diseases: A Narrative Review. Microorganisms, 11(5), 1269. https://doi.org/10.3390/microorganisms11051269 DOI: https://doi.org/10.3390/microorganisms11051269

Nagakubo, D., & Kaibori, Y. (2023). Oral Microbiota: The Influences and Interactions of Saliva, IgA, and Dietary Factors in Health and Disease. Microorganisms, 11(9), 2307. https://doi.org/10.3390/microorganisms11092307 DOI: https://doi.org/10.3390/microorganisms11092307

Nagarajappa, S., Bathija, P., Mishra, P., Bansal, V., Gupta, S., & Sontakke, S. (2018). Antibacterial and Antifungal Activity of Neem and Clove Extract Against S. mutans and C. albicans-An invitroStudy. World Journal of Pharmaceutical Research, 7(5), 1484-93.

Padminee, K., Anusha, R., Lakshmi, K., & Kumar, P. D. M. (2020). Geographic disproportions in dental workforce distribution and its impact on oral disease burden: An Indian perspective. SRM Journal of Research in Dental Sciences, 11(2), 76-81. DOI: https://doi.org/10.4103/srmjrds.srmjrds_79_19

Pasupuleti, M. K., Nagate, R. R., Alqahtani, S. M., Penmetsa, G. S., Gottumukkala, S. N. V. S., & Ramesh, K. S. V. (2023). Role of Medicinal Herbs in Periodontal Therapy: A Systematic Review. Journal of International Society of Preventive & Community Dentistry, 13(1), 9–16. https://doi.org/10.4103/ jispcd.JISPCD_210_22 DOI: https://doi.org/10.4103/jispcd.JISPCD_210_22

Paul, O., Arora, P., Mayer, M., & Chatterjee, S. (2021). Inflammation in periodontal disease: possible link to vascular disease. Frontiers in physiology, 11, 609614. DOI: https://doi.org/10.3389/fphys.2020.609614

Prabuseenivasan, S., Jayakumar, M., & Ignacimuthu, S. (2006). In vitro antibacterial activity of some plant essential oils. BMC complementary and alternative medicine, 6, 39. https://doi.org/10.1186/1472-6882-6-39 DOI: https://doi.org/10.1186/1472-6882-6-39

Rajhans, N. S., Kohad, R. M., Chaudhari, V. G., & Mhaske, N. H. (2011). A clinical study of the relationship between diabetes mellitus and periodontal disease. Journal of Indian Society of Periodontology, 15(4), 388–392. https://doi.org/10.4103/0972-124X.92576 DOI: https://doi.org/10.4103/0972-124X.92576

Rathee, M., & Jain, P. (2023).Gingivitis.In StatPearls. StatPearls Publishing.

Roller, S. (Ed.). (2003). Natural antimicrobials for the minimal processing of foods. Wood head publishing. DOI: https://doi.org/10.1533/9781855737037

Saigal, S., Bhargava, A., Mehra, S. K., & Dakwala, F. (2011). Identification of Candida albicans by using different culture medias and its association in potentially malignant and malignant lesions. Contemporary clinical dentistry, 2(3), 188–193. https://doi.org/10.4103/0976-237X.86454 DOI: https://doi.org/10.4103/0976-237X.86454

Shamsudin, N. F., Ahmed, Q. U., Mahmood, S., Ali Shah, S. A., Khatib, A., et al. (2022). Antibacterial Effects of Flavonoids and Their Structure-Activity Relationship Study: A Comparative Interpretation. Molecules (Basel, Switzerland), 27(4), 1149. https://doi.org/10.3390/molecules27041149 DOI: https://doi.org/10.3390/molecules27041149

Subramaniam, S.K., Siswomihardjo, W., & Sunarintyas, S. (2005). The effect of different concentrations of Neem (Azadiractha indica) leaves extract on the inhibition of Streptococcus mutans (In vitro). Dental Journal: Majalah Kedokteran Gigi, 38, 176-179. DOI: https://doi.org/10.20473/j.djmkg.v38.i4.p176-179

Taheri, J. B., Azimi, S., Rafieian, N., & Zanjani, H. A. (2011). Herbs in dentistry. International Dental Journal, 61(6), 287–296. https://doi.org/10.1111/j.1875-595X.2011.00064.x DOI: https://doi.org/10.1111/j.1875-595X.2011.00064.x

Van Dyke, T. E., & Sheilesh, D. (2005). Risk factors for periodontitis. Journal of the International Academy of Periodontology, 7(1), 3–7.

Van Dyke, T. E., Bartold, P. M., & Reynolds, E. C. (2020). The Nexus Between Periodontal Inflammation and Dysbiosis. Frontiers in immunology, 11, 511. https://doi.org/10.3389/fimmu.2020.00511 DOI: https://doi.org/10.3389/fimmu.2020.00511

Vaou, N., Stavropoulou, E., Voidarou, C., Tsakris, Z., Rozos, G., Tsigalou, C., & Bezirtzoglou, E. (2022). Interactions between medical plant-derived bioactive compounds: Focus on antimicrobial combination effects. Antibiotics, 11(8), 1014. DOI: https://doi.org/10.3390/antibiotics11081014

Wilson, C., Lukowicz, R., Merchant, S., Valquier-Flynn, H., Caballero, J., et al. (2017). Quantitative and Qualitative Assessment Methods for Biofilm Growth: A Mini-review. Journal of engineering and technology, 6(4), 1.25.

Wolinsky, L. E., Mania, S., Nachnani, S., & Ling, S. (1996). The inhibiting effect of aqueous Azadirachta indica (Neem) extract upon bacterial properties influencing in vitro plaque formation. Journal of Dental Research, 75(2), 816–822. https://doi.org/10.1177/00220345960750021301. DOI: https://doi.org/10.1177/00220345960750021301

Wylie, M. R., & Merrell, D. S. (2022). The Antimicrobial Potential of the Neem Tree Azadirachta indica. Frontiers in pharmacology,

, 891535. https://doi.org/10.3389/fphar.2022.891535 DOI: https://doi.org/10.3389/fphar.2022.891535

Zhang, Y., Wang, Y., Zhu, X., Cao, P., Wei, S., & Lu, Y. (2017). Antibacterial and antibiofilm activities of eugenol from essential oil of Syzygium aromaticum (L.) Merr.& L. M. Perry (clove) leaf against periodontal pathogen Porphyromonas gingivalis. Microbial pathogenesis, 113, 396–402. https://doi.org/10.1016/ j.micpath.2017.10.054 DOI: https://doi.org/10.1016/j.micpath.2017.10.054

Zhu, L., Tang, Z., Hu, R., Gu, M., & Yang, Y. (2023). Ageing and Inflammation: What Happens in Periodontium?. Bioengineering (Basel, Switzerland), 10(11), 1274. https://doi.org/10.3390/ bioengineering10111274 DOI: https://doi.org/10.3390/bioengineering10111274

Downloads

Published

2024-07-15

How to Cite

Bhattacharya, T., Saha, R. P., Majumder, R., & Ghosh, S. (2024). Investigating the antimicrobial activity of neem and clove extract on biofilm-producing oral microflora. Journal of Experimental Biology and Agricultural Sciences, 12(3), 511–521. https://doi.org/10.18006/2024.12(3).511.521

Issue

Section

PROCEEDING OF BIONEXT-2023_RESEARCH ARTICLES