Project Proposal

(Rough Protocol) Metal ion toxicity test 1. Collect the soil samples from metal enriched soil such as from in and around the mining areas. 2. The sample will be brought to the lab and serial dilution using sterile distilled water is done. 3. The sample is plated on the nutrient agar medium and PDA kept for incubation. 4. Single colony isolation was done to obtain pure strains. 5. The bacterial strain is identified and characterized. (The stain of the bacteria was reconfirmed by the microbiologist). 6. The growth curve of the specific strain is studied. 7. The resistance to anti-bacterial agents was tested and toxicity level was determined. 8. This strain of bacteria was subjected to of various toxic metals, such as Cd, Pb, Co, Zn, Hg, Ag, selenite, tellurite and uranyl. 9. The range of the resistance level of the strain of the bacteria was determined. 10. The tolerance of the bacteria to the metal ion is due to its capacity to reduce it to its elemental state which neutralizes its toxic effects. 11. Transmission electron microscopy and energy dispersive X-ray analysis was done to analyse the intracellular concentrations and state of these metals present there. 12. The percentage of metal ion uptake was determined based on the biomass concentration of each cell. 13. The change in the amino acid pool in the intracellular matrix with respect to the type and concentration of the metal ion intake was determined by analysis of soluble thiols using............ 14. To verify the hypothesis of over expression of efflux systems to get rid of drugs and heavy metals, analyse and localize the elemental composition of bacteria grown in the presence of tellurite and selenite, by using Energy Dispersive X-ray Spectroscopy (EDX) in conjunction with Transmission Electron Microscopy (TEM) or Environmental Scanning Electron Microscopy (ESEM). 15. The chemical properties essential for bactericidal activity by certain minerals mixture were probed by testing antibacterial activity in the presence of metal chelators like EDTA or desferrioxamine, the hydroxyl radical scavenger, thiourea, and varying pH levels.(NMR) 16. Effect of the acidic environment of the hydrated minerals on antibacterial activity is determined. 17. Changes in the above analysis when the sample was treated with metal oxide nano-particles (FeO2).

Oral Biofilm Architecture on Natural Teeth

Abstract: Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and accessibility. Despite descriptions of initial plaque formation on the tooth surface, studies on mature plaque and plaque structure below the gum are limited to landmark studies from the 1970s, without appreciating the breadth of microbial diversity in the plaque. We used fluorescent in situ hybridization to localize in vivo the most abundant species from different phyla and species associated with periodontitis on seven embedded teeth obtained from four different subjects. The data showed convincingly the dominance of Actinomyces sp., Tannerella forsythia, Fusobacterium nucleatum, Spirochaetes, and Synergistetes in subgingival plaque. The latter proved to be new with a possibly important role in hostpathogen interaction due to its localization in close proximity to immune cells. The present study identified for the first time in vivo that Lactobacillus sp. are the central cells of bacterial aggregates in subgingival plaque, and that Streptococcus sp. and the yeast Candida albicans form corncob structures in supragingival plaque. Finally, periodontal pathogens colonize already formed biofilms and form microcolonies therein. These in vivo observations on oral biofilms provide a clear vision on biofilm architecture and the spatial distribution of predominant species.