The effect of a homoeopathic and herbal mouthwash formulation on the growth of Streptococcus salivarius and Fusobacterium nucleatum in vitro

Abstract:There are a few hundred different types of bacterial species within the oral cavity which amount to more than several 100 millions in number within dental plaque. Some bacteria are commensal and others play a key role in the progression of periodontal disease. Periodontal disease can manifest as gingivitis which is seen as infection of the gingival tissue, or periodontitis which results in loss of collagen attachment of tooth to bone and loss of bone tissue. Streptococcus salivarius is the principal commensal bacterium in the oral cavity of humans. Fusobacterium nucleatum is commonly found in the dental plaque of humans and is frequently associated with periodontal disease. The mouthwash formulation is sea salt based together with homoeopathic remedies and herbal extracts. It is indicated for the treatment of sensitive irritated gums, halitosis, sore throat and mouth ulcers. The aim of this study was to determine the in vitro effect of a homoeopathic and herbal mouthwash formulation on Streptococcus salivarius and Fusobacterium nucleatum. This study forms part of a three part in vitro study to determine the effect of the mouth wash formulation. The effect was evaluated by measuring zones of inhibition around discs impregnated with the mouthwash solution, saline solution and distilled water. Lypholized ATCC 25586 Fusobacterium nucleatum and ATCC 13419 Streptococcus salivarius were obtained from Quantum Biotechnologies. Fusobacterium nucleatum was cultured on chocolate blood agar and nutrient agar and incubated at 35 ᵒC in 5-7% CO2 for 48 hours. Streptococcus salivarius was cultured on Tryptic Soy Agar (TSA) agar with 5% sheep blood and incubated at 35 ᵒC in 5-7% CO2 for 48 hours. Agar plates were refrigerated at 2-8 ᵒC for two weeks and used as stock cultures. In the Kirby-bauer disk diffusion method, agar plates incubated with Fusobacterium nucleatum and Streptococcus salivarius respectively were streaked with the mouth wash solution, and the control substances distilled water and saline solution respectively. The agar plates were incubated at 35 ᵒC in 5-7% CO2 for 48 hours. In Adaptation experiment one, a five times, and two times strength mouth wash solution was prepared and tested following the Kirby-bauer streaking method. In the Adaptation experiment two, a volume of 100 μl of one time, two times and five times strength mouth wash solution was mixed with 10 μl cultures of Streptococcus salivarius and Fusobacterium nucleatum in a 1ml eppendorf tube. The mixture was incubated for 5 minutes at room temperature and streaked on TSA agar with 5% sheep blood and nutrient agar plates respectively, agar plates were incubated at 35 ᵒC in 5-7% CO2 for 48 hours