ANTIMICROBIAL ACTIVITY OF ALCHORNEA CORDIFOLIA AGAINST EXTENDED SPECTRUM BETA LACTAMASE PRODUCING ESCHERICHIA COLI AND KLEBISELLA PNEUMONIAE



INTRODUCTION
The history of the use of herbs in the management of diseases dates back to the time of the early man (Sofowora, 1982 and Kafara, 1994). In herbal medicine, herbs / plants are being used in their unaltered form for the treatment of disease. (Physiological society of Nigeria, 2009). The WHO estimates that nearly 70% of the world population depend on traditional medicine, especially medicinal plants, for their primary health needs (Ansah. et al, 2011).

Alchornea cordifolia, a medium-sized shrubby tree is one of such herbs. It belongs to the subfamily Acalypholdeae and family Euphorbiaceae or spurge family. It is sometimes scrambling bushy, perenial shrub or small tree, up to 4m high reproducing from seeds. (Osadebe. et al, 2012). Its common name includes Poruru, Poruro, Purosa, macochichua and Nianda. The English name is Christmas bush, and Efik name is Mbom (Physiological society of Nigeria, 2009). It is also known as Ewe Ipa, Ubobo and Bambami in Nigeria. (Adeshina. et al, 2012).
It is widely distributed from Senegal east to Kenya, Tanzania, South and Central Africa to Angola it is cultivated in DR congo for its medicinal use (Mavar-Mange. et al, 2004), and geographically distributed in secondary forest usually near water, moist or marshy places. (Adeshina. et al, 2012).
Alchornea cordifolia is commonly used as a medicinal plant throughout its area of distribution. The leaves are mostly used, but also the stem bark, stem pith, leafy stems, root bark, roots and fruits enter into local medicine. The leaves or leafy stems, are chewed fresh taken for their sedative and antispasmodic problems including sore throat, cough and bronchitis. Genital-urinary problem including venereal diseases and female sterility. It can also be used to treat intestinal problems including gastric-ulcers, diarrhoea, amoebic dysentery and worms. They are also taken as blood purifier, as a tonic and to treat anaemia and epilepsy. In Nigeria a decoction of bruised fruit is taken to prevent miscarriage.
It has been found to have anti-inflammatory, antibacterial and analgesic properties (Cesario, 1993). The phytochemical properties include alchorneine, anthranilic acid, gentisnic acid, iso-alchorneine, yohimbine and alkaloids (Duke and Vasquez, 1994).
A majority of Nigerian population still rely on Traditional Medicine. Available evidence suggests that some herbal remedies and traditional therapeutic regimes are efficacious and affordable (TMP, 2007).
The aqueous and ethanol extracts from Alchornea cordifolia were found to inhibit to growth of some negative bacteria and E. coli and K. pneumoniae being gram-negative bacteria were believed to be inhibited (Ajao. et al, 1985).

PROPERTIES
The roots and the stem bark contain trepenoids, steroid, glycosides, flavonoids (2-3%), tannis (about 10%), saporins, carbohydrates and the imidazopyrimidine alkaloids. The leaves also contain a range of hydroxybenzoic acid, gallic acid, ethyl ester, gentisic acid, anthranilic acid (vitamin L1) and protocatechuic acid (Mavar-Manga. et al, 2007)

AIM
To detect the sensitivity of Alchornea cordifolia stem, leaves, root and bark on the E. coli and K. Pneumoniae as an extended spectrum beta lactamase producing organisms.

OBJECTIVES
To shown that Alchornea cordifolia is a good herb for infections and diseases caused by E. coli and K. Pneumoniae.
To show that Alchornea cordifolia is also a good herb for infections caused by E. coli and K. pneumoniae producing extended spectrum beta lactamase enzyme.
To show that K. Pneumoniae. and E. coli can be present in a healthy individual without causing diarrhea.

MATERIALS AND METHODS
Collection of Alchornea cordifolia leaf, stem, root, and bark
-           Preparation of their extracts and its derived fractions
-           Phytochemical screening
-           Soxhlet extractor
-           dessicator
-           Extraction of the secondary metabolites
-           Collection of feaces from health staffs of a fast food joint in Abakaliki. To isolate E. coli and K. Pneumoniaes in extended spectrum beta lactamase enzyme.
        
COLLECTION OF ALCHORNEA CORDIFOLIA PARTS
Stem, roots, barks and leaves of Alchornea cordifolia  will be collected from Abakaliki.                 
   
IDENENTIFICATION AND AUTHENTICATION
The parts of alchornea cordifolia parts will be identified and authenticed in near research institute.

EXTRACTION OF THE PARTS WITH ETHANOL, METHANOL, ACETONE HOT WATER AND COLD WATER
After drying of the parts,they will be reduced to powderd form. The active parts will be extracted with ethanol, methanol,acetone,hot and cold water with a soxhlrt extractor and kept in a dessicator.

DETERMINATIONM OF THE ACTIVITY OF THE PLANT EXTRACTS ON EXTENDED SPECTRUM BETA LACTAMASE (ESBL) PRODUCING BACTERIA.
The reaction of the plants extracts on the isolated ESBL bacteria organisms (E.coli and K.pneumoniae) will be determined.

ISOLATION AND CHARACTERIZATION OF THE BACTERIA PRODUCING ESBL ENZYMES.
E.coli forms circular convex, smooth colonies with distinct edges, ferment lactose, produce Shemolysis on blood agar,motile, oxidase and citrate negative, positive for indole, lysine decarboxylase and manitol fermtation whereas K.pneumoniae produce large and regular polysaccharide capsules,colonies are large and muciod,tent to coalesce with prolonged incubation, lack motility, positive for lysine decarboxylase and citrate.

SUSCEPTIBILITY TESTING
It will be carried out according to McFarland scales (0.5 McFarland is about 106  cfu/ml ) and a sterile cork borer will be used to bore equidistant cups on the agar plates.

DETERMINATION OF MINIMIUM INHIBITORY CONCENTRATION (M.I.C).
Sterile Mueller-Hinton agar plates will be prepared using double dilution method. The different parts of solidified Alchornea cordifolia admixture plates will be inoculated with 2.0ml of standardized 18  hour culture test organism. The inocula will be allowed to diffuse into the test agar plates for 30 min and incubated at 37oC for 18 hour. The lowest concentration of the extracts in the test agar plates that showed no growth will be considered as the M.I.C of the extract against the test organism.

DETERMINATION OF MINIMIUM BACTERIA CONCENTRATION (M.B.C).
This will be carried out by inoculating the concentration of the extract/fraction sterile nutrient broth test-tubes containing inactivating agents 3% v/v TWEEN 80. the test-tube will be incubated at 37oC for 24 hours after which they will be examined for presence or absence of growth.

SCREENING FOR THE PHYTOCHEMICAL COMPOTENT OF PLANT EXTRACT
The phytochemical analysis of the active parts will be determined to detect the presence of flavonoids,trepenoids, steroid, glycosides, tannis, saporins,carbohydrates imidazopysionidine alkaloids, gallic acid, ethyl ester, gentisic acid, hydroxybenzoic acids, anthranilic acid and protocatechuic acid.

REFERENCES
Ajao A.O, Shonukan O. and Femi-Onadeko B.(1985).Antibacterial effect of aqueous and alcohol extracts of spondias mombin and Alchornea cordifolia. Two local antimicrobial remedies. Vol.23 (2):67-72.

Caseerio, A. (1993).  Screeening of plants used in argentine folk medicine for antimicrobial activity.Journal of Enthnopharmacology,Vol 139:119-128.

Charles Ansah, Emmanuel Oppong and Eric Woode. (2011). Subacute Oral Toxicity assessment of Alchornea cordifolia (schumach and Thonn). Vol. 10 (5): 587-594.

Duke J Vasquez,R (1994). Amazonian Entnobotanical Dictionary,Florida.CRC Press Incorporated.vol 112:101-102.

G.O. Adeshina, O.F. Kunle, J.A. Onaolapo, J.O. Ehinmidu and L.E. Odama (2012) Phytochemical and Antibacteria I studies of the hexane  Extract Phytochemicals as Nutraceuticals – global approaches to their Role in Nutrition  and Health, Dr. venketeshwer Rao (Ed), ISBN Available from
http://www.intechopen.com/books/phybochemical-as-nutraceuticals-global-approaches to their role of Alchornea cordifolia leaf.

Mavar-Manga H, Lejoly J,Quetin-Leclercq J. and Schmelzer,G.H (2007). Alchornea cordifolia  (Schumach and Thonn.) Mull Arg.PROTA.

Mavar-Manga H,Brick O,D Marie D.E.P and Quetin-Leclercq J.(2004). Journal of Enthnopharmacology. Vol 92(2-3):209-214.

Nigerian Journal of Physiological Sciences (2009). Vol 24(2):149-151. Physiological Society of Nigeria.

Patience O. Osadebe, Fetus BC Okoye, Philip F. Uzor, Nneka R. Nnamani, Ijeoma O.E. Adide and Nkemakonam C. Obiano (2012). Phytochemical analysis, hepatoprotective and antioxidant activity of alcrornea cordifolia methanol leaf extract on carbon tetrachloride induced hepatic damage in rats. 289-293.

TMP (2007).Traditional Medicine Policy for Nigerian. Pp. 66.
Zuleikha N. A, Omonkhelin O. J. Buniyamin AA, Paul 10, (2010) Evaluation of the oxytocic activity of the ethanol extract of the roots of Alchornea cordifolia. In green Pharm. 4:247-250. The extracts from leaves of A. Cordifolia were found to inhibit the growth of bacteria such as E. coli 


DEPARTMENT:                 APPLIED MICROBIOLOGY
FACULTY:                           BIOLOGICAL SCIENCES




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