title: Antibacterial and anti-adherence effects of a plant extract mixture (PEM) and its individual constituent extracts (Psidium sp.,Mangifera sp., andMentha sp.) on single- and dual-species biofilms.
abstract: BACKGROUND
  Plant extracts mixture (PEM) and its individual constituent plant extracts(Psidiumsp.,Mangiferasp.,Menthasp.) are known to have an anti-adhering effect towards oral bacteria in the single-species biofilm. To date, the adhering ability of the early and late plaque colonisers (Streptococcus sanguinisandStreptococcus mutans) to PEM-treated experimental pellicle have not been investigated in dual-species biofilms.
  
  
  METHODS
  Fresh leaves of these plants were used in the preparation of the respective aqueous extract decoctions. The minimum inhibitory concentration (MIC) of the extracts towardsS. sanguinisATCC BAA-1455 andS. mutansATCC 25175 was determined using a two-fold serial microdilution method. The sum of fractional inhibitory concentration (ΣFIC) index of PEM and its constituent plant extracts was calculated using the MIC values of the plants. The minimum bactericidal concentration (MBC) of the plant extracts was also determined. The anti-adherence effect of the plant extracts (individually and mixed) was carried out by developing simulatedS. sanguinisandS. mutansrespectively in single- and dual-species of biofilms in the Nordini's Artificial Mouth (NAM) model system in which the experimental pellicle was pretreated with the plant extract before bacterial inoculation. The bacterial population in the respective biofilms was quantified using ten-fold serial dilutions method and expressed as colony forming unit per ml (CFU/ml). The bacterial population was also viewed using Scanning Electron Microscope (SEM). All experiments were done in triplicate.
  
  
  RESULTS
  The PEM compared with its respective constituent plants showed the lowest MIC towardsS. sanguinis(3.81 mg/ml) andS. mutans(1.91 mg/ml) and exhibited a synergistic effect. ThePsidiumsp. (15.24 mg/ml) and, PEM andPsidiumsp. (30.48 mg/ml) showed the lowest MBC towardsS. sanguinisandS. mutansrespectively. The anti-adherence effect of the PEM and its respective constituent plants (exceptPsidiumsp.) was different for the two bacteria in the single-species biofilm. In the dual-species biofilms, PEM demonstrated similar anti-adherence effect towardsS. sanguinisandS. mutans. The proportions of the bacterial population viewed under SEM appeared to be in agreement with the quantified population.
  
  
  CONCLUSIONS
  The combination of the active constituents of the individual plant extracts in PEM may contribute to its low MIC giving rise to the synergistic effect. The different anti-adherence effect towardsS. sanguinisandS. mutansin both single- and dual-species biofilms could be due to the different proportion of the active constituents of the extracts and the interaction between different bacteria. The better adhering ability ofS. sanguinistowards the PEM-treated pellicle when present together withS. mutansin the dual-species biofilms may suggest the potential of PEM in controlling the balance between the early and late colonisers in biofilms.
authors.affiliation: Department of Oral Biology and Biomedical Sciences, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia; Department of Clinical Oral Biology, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Zaleha
authors.initials: Z
authors.lastname: Shafiei
authors.affiliation: Department of Oral Biology and Biomedical Sciences, Faculty of Dentistry, University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Zubaidah
authors.initials: Z
authors.lastname: Haji Abdul Rahim
authors.affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya , Kuala Lumpur , Malaysia.
authors.forename: Koshy
authors.initials: K
authors.lastname: Philip
authors.affiliation: Centre for Pre-U Studies, UCSI University , Kuala Lumpur , Malaysia.
authors.forename: Nalina
authors.initials: N
authors.lastname: Thurairajah
keywords: NAM system; Scanning Electron Microscopy; Streptococcus mutans; Streptococcus sanguinis