1. Screening of 40 Plant extracts for antimicrobial activity against Mycobacterium tuberculosis Richin John Koshy Supervisor: Maria Lerm

2. Overview Background Hypothesis/Aim Materials and methods Results Conclusion

4. Morphology Mycobacteria are small rod shaped aerobic bacilli Mycobacteria are present in the environment, soil and water Mycobacteria have high concentration of lipids and wax in the cell wall making them resistant to standard staining techniques Mtb grow quite slowly and colonies do not appear on solid media up to 2.5 -5 week Media which are used in growth of mycobacteria are selective media, semisynthetic media and broth media www.lookfordiagnosis.com

5. Pulmonary TB

6. commons.wikimedia.org

7. Treatment of TB niaid.nih.gov

8. Drug Development Screening of Chinese herbal medicine led to introduction of artemisinin from the leaves of Artemisia annua which is used worldwide for treatment against malaria At present, there are eight to ten drugs with therapies lasting to 18-24 months of which there are only four drugs which are actually developed to treat TB Shifting the screening strategy from a single enzyme targets to a whole bacterial cell level have been showing successful results as it has a holistic approach and only gives hits that can enter the bacterium through the cell wall

9. Hypothesis Whole cell screening method is a more effective way than molecular/chemical approaches to identify new, anti-mycobacterial drug candidates/lead compounds Traditionally used plants are a valuable source for the discovery of modern drugs

10. Aim To screen 40plant extracts prepared from plants used traditionally to treat tuberculosis and other infections in Sudan for possible antimicrobial activity against Mtb

11. Materials and Methods

12. Results Effect of DMSO on plant extracts DMSO at concentration of 10 %used along with plant extracts as treatments for reduction of antimycobacterial growth showing higher pattern of inhibition and a reduction of inhibition seen with DMSO concentration of 1 % and 0.1 %

13. Anti-mycobacterial screening of plant extracts against avirulent strain of Mtb H37Ra (1)

14. Anti-mycobacterial screening of plant extracts against avirulent strain of Mtb H37Ra (2) Screening of 40 plant extracts by analysis of mycobacterial growth by luminometry and measuring cell viability showing a pattern of inhibition in plant extracts A10, A12, A20, A22, A28, A29, A30, A34

15. Confirming screening of plant extracts showing inhibition Student paired t test was done assuming Gaussian distribution which show statistically insignificant hits

16. Detecting changes in mycobacterial growth by CFU plating

17. Determination of pH

19. 40 extracts of selected medicinal plants were analysed for antimycobacterial activities which yielded 5 extracts to have distinct antimycobacterial properties

20. Further studies were carried out in order to measure colonies in the presence of selected medicinal extracts showing inhibition of bacterial growth, thereby ruling out inhibition due to luciferase

22. I also express my gratitude to Johanna Raffetseder, Marie Larsson, ElsjePienaarand Daniel Eklundfor introducing me to most of the methods iused in this project and more and also for always being there to answer numerous questions I had.