in vitro antimicrobial activity of aloe turkanensis , an ethnomedical herbal remedy used in...

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

Introduction

Five common speciesAT…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 malariaWoundsStomachachesPainRingworms 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 methodsCollection of the Plant MaterialWhole 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

Malaria 10 WP Clinical signs Boiled leaf extract when dry

Oral

Wounds 9 Leaf Clinical signs Aloe juice Topical

Pain 8 Whole Clinical signs Boiling when dry OralStomach ache 8 Leaf Clinical signs Boiling when dry Oral

Tiredness 9 Leaf Clinical signs Boiling when dry OralDetoxify 6 Whole Clinical signs Boiling when dry Oral Laxative 10 Leaf Clinical signs Boiling when fresh Oral

Emetic 3 Leaf Clinical signs Boiling when fresh Oral

Cosmetic 10 Leaf Clinical signs Freshly harvested sap & gel Topical

Ringworms 7 Leaf Clinical signs Freshly harvested sap Topical

Health Drink 10 Leaf & roots Clinical signs Gel Oral

Removal RAB in cows

2 Leaf Clinical signs Sap Intrauterine

Chicken disease 2 Leaf Clinical signs Freshly harvested sap Drinking water

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

Preparation of plant samples Washed, chopped dried hot air oven 800C 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 370C …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…-veBenzyl penicillin, Gentamycin and Amphotericin B …+VE

Each tube containing the test organism and plant extract was the incubated for 24 hours at 370C…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 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.

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

Results/Discussion

Test material Extract Part used P. aeroginosa E. coli S. aureus B. cereus C. albinans

A. turkanensis Aqueous WP - - <3.125 200 -

Methanol WP 200 - 100 100 -

Ag Sap 125 500 62.5 62.5 -

Met Sap - - 62.5 62.5 -

Benzyl penicillin - - - 0.625 0.625

Gentamycin - - 0.0049 0.0049 - - -

Amphotericin B - - - - - - 0.0125

ConclusionBased 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 PhytochemistryWhat is the significance of the noted

biological activity?...

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