In vitro Antimicrobial Activity of Aloe turkanensis , an Ethnomedical Herbal Remedy Used in Turkana ,

1. In vitro Antimicrobial Activity of Aloe turkanensis, an Ethnomedical Herbal Remedy Used in Turkana, Z. M. RUKENYA1, J. M. MBARIA1, P.M. MBAABU2, S. G. KIAMA2 R.O.ONZAGO1

2. Introduction • Five common species • AT…Important ethnomedical remedy.

3. Sap used for wound management & eye diseases. • Boiled roots juice in soup induce vomiting relieve persistent headaches. • The roots are used to flavour beer (Bosch, 2006). • Natira community aloe-working group herbalist • Treat malaria • Wounds • Stomachaches • Pain • Ringworms • Poultry diseases.

4. Justification • Despite wide usage, there is inadequate technical specifications on quality, safety and efficacy (no available microbial load standards---KEBS). Culture on BA

5. Scanty literature on AT…Probably following presidential degree in 1986 … • Study done by Schmelzer (Protabase), et al 2010…Knowledge gaps in quality control standards for exudates of Aloe secundifloraand Aloe turkanensis • Repercussions: • Hindrance to international trade • Why Aloe turkanensis(land rehabilitation, drug discovery, public health, livelihoods-value chain) • Good news…C & M of Aloes L.N 403 2007 gazette…80% ASAL..CBS 1999.

6. Hypothesis • Aloe-working group herbalists recognize that wounds, eye infections, fungal infections, stomach upsets and skin infections involve pathogenic micro-organisms and that the plant Aloeturkanensis has metabolites capable of inhibiting or killing these microbes without causing adverse effects on the herbal consumer. Am 95% safe if consumed

7. Objectives • Determine the bioactivity of the plant’s extract (both naturally occurring and cultivated) on the growth of Bacillus cereus, Staphylococcus aureus, Pseudomonas aeroginosa, Escherichia coli, Candida albicansand laboratory culturedArtemiasalinaleach

8. Materials and methods Collection of the Plant Material • Whole plant was harvested from Natira community aloe garden…attended mainly by local women. Kakuma town Natira sub-location • Collection was done during a dry season

9. Aided by 10 people from Natira community Aloe-working group herbalists who identified the plant by the local name and voluntarily provided information on uses Aloe turkanensis as medicinal plant

10. A plant specimen was identified at KFRI, stored and young ones propagated

11. Preparation of plant samples • Washed, chopped dried hot air oven 800C 72 hours then milled to powder using a laboratory grinder. Sap screened.

12. Extraction process • Cold maceration • Rota-Evaporation to dryness • Lyophilized to powder using Christ Beta lyophilizer. The aqueous extract yielded 13.32g while methanolic 14.96g. Stored in a refrigerator pending further analysis.

13. Testing for antimicrobial activity of the extracts • Broth dilution assays on 4 standard cultures & 1 clinical isolate • Escherichia coli (ATCC 25922), Pseudomonas aerugenosa(ATCC 27853), Staphylococcus aureus (ATCC25923) & Bacillus cereus (ATCC 11778) while human clinical isolate of Candida albicans • Minimal inhibitory concentrations (MIC) determined. • Reference bacterial obtained from cooked meat media • Subcultured in blood agar (Oxoid®) for 24 hours at 370C …except for Candida albicans incubated at rt.

14. Using a sterile loop, colonies picked & put into 3ml sterile physiological buffer saline (PBS). • Serial dilution…Macfarland opacity No. 6 • Stock solution of the plant extracts • 4 grams of powder dissolved 10ml of MHB to make a concentration of 400mg/ml…then two fold serial dilutions made in sterile Muller Hinton Broth (MHB) using culture tubes. • One ml of the test organisms’ suspension dispensed into the culture tubes containing the plant extracts. • Controls • 1ml of the test organisms’ suspension in sterile MHB…-ve • Benzyl penicillin, Gentamycin and Amphotericin B …+VE • Each tube containing the test organism and plant extract was the incubated for 24 hours at 370C…Candida albicansRT 48 hrs.

15. The inhibition of test organisms was evaluated by culturing 1ml of the suspension into Muller Hinton Agar for 24hour at 370C for bacteria and for 48 hours at room temperature for Candida albicans. Brine Shrimp Lethality Test: • Lethality testing was done according to McLaughlin (1991). Three dilutions were prepared by transferring 500µl, 50µl and 5µl of plant extract to a set of five graduated tubes. • Ten shrimp were transferred into each of the vial using Pasteur pipette and marine salt added up to 5ml mark to make dilution of 1000µg/ml, 100µg/ml and 10µg/ml. After 24 hours exposure, live nauplii were counted.

16. Results/Discussion Table 3: Minimum Inhibitory Concentration (MIC mg/ml) of Aloe turkanensis extracts after incubation with microbial cultures

17. Conclusion • Based on interviews conducted in the field and the subsequent in vitro laboratory testing of the sampled plant, it was concluded that the plant has metabolites that inhibit the growth of some bacterial organisms and is a relatively safe herbal remedy when used for human consumption. It is probably due to the antibacterial activity and safety of Aloe turkanensis that Natira community Aloe-working group herbalists use the plant products for treatment of wounds, eye infections , skin infections, stomach upsets, livestock diseases and as cosmetics.

18. Pending work • Phytochemistry • What is the significance of the noted biological activity?...

19. ACKNOWLEDGEMENTSRISE-AFNNET (SIG)UONAUDIENCETHANK YOU