Presentation on theme: "Z. M. RUKENYA 1, J. M. MBARIA 1, P.M. MBAABU 2, S. G. KIAMA 2 R.O.ONZAGO 1."— Presentation transcript:
Z. M. RUKENYA 1, J. M. MBARIA 1, P.M. MBAABU 2, S. G. KIAMA 2 R.O.ONZAGO 1
Introduction Five common species AT…Important ethnomedical remedy.
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.
Justification Despite wide usage, there is inadequate technical specifications on quality, safety and efficacy (no available microbial load standards- --KEBS). Culture on BA
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 secundiflora and 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.
Aloe-working group herbalists recognize that wounds, eye infections, fungal infections, stomach upsets and skin infections involve pathogenic micro-organisms and that the plant Aloe turkanensis has metabolites capable of inhibiting or killing these microbes without causing adverse effects on the herbal consumer. Hypothesis Am 95% safe if consumed
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 albicans and laboratory cultured Artemia salina leach Objectives
Materials and methods Collection of the Plant Material Whole plant was harvested from Natira community aloe garden…attended mainly by local women. Natira sub-location Kakuma town Collection was done during a dry season
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 AILMENT No. Of people who have used within the last 10 years Part used Method of disease diagnosis Method of reconstituting the medicine Route of administration Malaria10WPClinical signsBoiled leaf extract when dry Oral Wounds9LeafClinical signsAloe juiceTopical Pain8WholeClinical signsBoiling when dryOral Stomach ache8LeafClinical signsBoiling when dryOral Tiredness9LeafClinical signsBoiling when dryOral Detoxify6WholeClinical signsBoiling when dryOral Laxative10LeafClinical signsBoiling when freshOral Emetic3LeafClinical signsBoiling when freshOral Cosmetic10LeafClinical signsFreshly harvested sap & gelTopical Ringworms7LeafClinical signsFreshly harvested sapTopical Health Drink10Leaf & rootsClinical signsGelOral Removal RAB in cows 2LeafClinical signsSapIntrauterine Chicken disease2LeafClinical signsFreshly harvested sapDrinking water
A plant specimen was identified at KFRI, stored and young ones propagated
Preparation of plant samples Washed, chopped dried hot air oven 80 0 C 72 hours then milled to powder using a laboratory grinder. Sap screened.
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. Extraction process
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 37 0 C …except for Candida albicans incubated at rt.
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 37 0 C…Candida albicans RT 48 hrs.
The inhibition of test organisms was evaluated by culturing 1ml of the suspension into Muller Hinton Agar for 24hour at 37 0 C 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.
Table 3: Minimum Inhibitory Concentration (MIC mg/ml) of Aloe turkanensis extracts after incubation with microbial cultures Results/Discussion Test materialExtractPart usedP. aeroginosaE. coliS. aureusB. cereusC. albinans A.turkanensisAqueousWP--<3.125200- MethanolWP200-100 - AgSap12550062.5 - MetSap--62.5 - Benzyl penicillin---0.625 Gentamycin--0.0049 --- Amphotericin B------0.0125
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.
Pending work Phytochemistry What is the significance of the noted biological activity?...
ACKNOWLEDGEMENTS RISE-AFNNET (SIG) UON AUDIENCE THANK YOU