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Sky Journal of Microbiology Research Vol. 3(4), pp.

041 - 045, August, 2015
Available online http://www.skyjournals.org/SJMR
ISSN 2315-876X ©2015 Sky Journals

Full Length Research Paper
Antibacterial activity of bitter leaf (Vernonia
amygdalina) soup on Staphylococcus aureus and
Escherichia coli)
Lovet T. Kigigha1* and Ebubechukwu Onyema2
Department of Biological Sciences, Niger Delta University Wilberforce Island, P. M. B., 71, Yenagoa, Bayelsa State,
Nigeria.
Accepted 25 July, 2015

The study was aimed at assessing the effect of cooking Egusi-soup on the antibacterial activity of bitter leaf
(Vernonia amygdalina). Extracts of the leaves of bitter-leaf plant were made using Ethanol, from Bitter-leaf soup
and using hot water. The antibacterial activity of the extracts was tested using two pathogenic hospital isolates
viz. Staphylococcus aureus and Escherichia coli. The antibiotic, Ampiclox (a broad- spectrum antibiotic in 1 mg
-1
ml concentration) was used as control. The test showed that all the three forms of extracts were inhibitory to
the two isolates in which the hot water extract indicated the highest inhibitory zone on E. coli and also with S.
aureus (P < 0.001). This was followed by the inhibitory effect of bitter leaf soup extract which was significantly
higher than the inhibitory effect of Ampiclox on E. coli (representing the enteric bacteria) (P = 0.025).
Interestingly, the effect of bitter leaf soup extract was also higher than the effect of Ampiclox on S. aureus
(representing superficial etiologic agents) indicating a probable positive effect on the health of probably
external and internal mucosa. The observed antibacterial effect of V. amygdalina on the selected bacterial
isolates appear to justify the traditional use of bitter leaf as a choice vegetable for African soup recipes and for
the cure of a number of human diseases (such as the treatment of gastroenteritis, fever and wound infections
etc).

Key words: Vernonia amygdalina, bitter leaf, antibiotic, Enterobacteriaceace, Escherichia coli, Staphylococcus aureus.

INTRODUCTION

Plants extracts are continuously being sort for as the roots of V. amygdalina have been used for gingivitis
effective and cheaper alternative sources of medication and toothache due to its proven antimicrobial activity.
all over the world especially in the developing countries. This study was designed to investigate the effect of
Vernonia amygdalina commonly called bitter leaf cooking Egusi soup (a major conduit for using bitter leaf
(because of its bitter taste) is consumed either as a in Nigeria and in Africa generally) on the antibacterial
vegetable for cooking African soups or the aqueous activity of the plant. This is because bitter leaf soup when
extracts could be drank as tonics for the treatment of consumed gives a great soothing, appetizing and
various illnesses (Imaga and Bamigbetan, 2003). The unusually satisfying after-effect.
bitterness is suspected to be due to factors such as the
presence of alkaloids, saponins, tannins and glycosides
which have been shown by various authors to be present LITERATURE REVIEW
in bitter leaf (Butler and Bailey, 1973; Ologunde et al.,
1992 and Bonsi et al., 1995). In the wild, chimpanzees Global use of medicinal plants
have been observed to ingest the leaves Vernonia plants
when suffering from parasitic infections. According to According to World Health Organization (WHO, 2007),
Huffman et al. (1993), medicinal plants are plants in which one or more of its
parts contain substances that can be used for therapeutic
purposes or which are precursors for the synthesis of
*Corresponding author. E-mail: lovet_k@yahoo.com. useful drugs. The local use of natural plants for primary

Latin V. Ibrahim et al. It was inferred that Africa.. magi. in the refrigerator. . would be of medicinal interest to ascertain the extent of Ewuro (in Yoruba) and Shiwaka (in Hausa). soup. the leaves could also be used for washing slime off fish and snail before cooking Preparation of plant extracts while roots and twigs could be used as chewing-stick. Previous research works on Vernonia amygdalina amygdalina include. It is also claimed refrigerator for use. palm oil. thus it Nigeria include: Kiriologbo (in Ijaw). The first batch Raserika. amygdalina. is a small ever green shrub that grows in tropical and antioxidants in the plant extract. Its mineral content (per gram) is and stored in the refrigerator for use. Other African soups. compounds for synthesizing analog drugs (Akerele. Though many nutritional and antimicrobial properties of V. amygdalina plants are consumed as food without an in-depth leaves. the soup. Onugbu (in Igbo).1 filter paper content 0·50%). onion. In many parts of west Africa. Selenium 8·2 µg. depreciation or otherwise of the known antibacterial The leaves are used as soup condiment and as activity of the plant when consumed in Egusi soup. Bitter leaf soup preparation was started with an initial 800 ml of water. In the same year. meat. contribution to health. The expressed 24 h. treatment of fever. (2009). their utilization through several amygdalina and Occimum gratissimum leaves on generations appear to justify their use (Ghani et al. This was then filtered using extract is used in treating skin infection such as Whatman No. selected food borne pathogens. 1992. the plants showed antibacterial activity on the entire test isolates. After Indication is that various studies have been carried out on cooking. pneumonia and pounded bitter leaf was filtered using 100 ml of boiled arthritis. 1994). 2002). (2009) carried out knowledge of their exact chemical composition and the assessment of the antibacterial activity of V. Staphylococcus aureus. Imaga and Bamigbetan (2013) carried out an in vivo biochemical assessment of aqueous extracts of V. vegetable after crushing and washing off using water to remove some of the bitterness (Mayhew and Penny. include: Ogbono and Okra soups.. This is because bitter with protenous ingredients (such as fresh or dry fish) in leaf soup when consumed gives a great soothing. Indication was that there was an improved functionality of the antioxidant system of the Vernonia amygdalina. in which bitter leaf is used MATERIALS AND METHODS apart from Egusi soup. Microbiol.. fresh pepper. salt. The 83·0%. Iron 4·71 µg and of 100 g pounded bitter leaf was used for preparing Egusi Zinc 1·13 µg. ii) The second batch of 100 g of to cure diabetes. The plant is mostly found in West Africa where the to body organs and tissues. iii) The third batch Phosphorus 61·55 µg.3 m in height with petiole leaf of aqueous extract of V. among others (Gill et al. Homoiona the leaf sample was rinsed three times with distilled and and Saffaf. All parts of the plant are pharmacologically suspected Samples of fresh plant of V..42 Sky. dry fish. researched on the American and Africa (Bibitha et al.. J. The plant is claimed to also exhibit deionized water. kidney disease and stomach The plant was used to prepare three different extracts: i) discomfort. amygdalina could be consumed as about 6 mm in diameter and elliptical in shape (Igile et al. 1998). a member of the Asteraceae test rats due to the probable effect of the phytochemicals family. Shigella dysentriae and Salmonella typhimurium in which 1993). soup thickener. protein 1·30% and ash extract was then filtered using what man No. Local names by which the plant is called in appetizing and unusually satisfying after-effect. Studies by Oboh and Masodje (2009) indicated water and stoppered with cotton wool covered with that V. Ingredients used alongside with V. It is a shrub of 1 . hiccups. 2003) as well as anti-tumourgenic properties was soaked in 100ml of ethanol which was stoppered (Izevbigie et al. Bacillus cereus. health remedies is a common practice in Asia. Fred et al. most used part is its leaves (called bitter leaf). amygdalina leaves were to be useful. Locally it is used in treating with cotton wool covered with aluminum foil and left for stomach ache (for immediate relief).1 filter paper and the extract stored in the ringworm. 2004). Both the roots and leaves are used in the bought at Amassoma market in Bayelsa state of Nigeria. rashes and eczema. Res. The aqueous and 1989). The leaves The present study was designed to investigate the are dark green in colour with a characteristic odour and effect of cooking Egusi soup (a major conduit for using when chewed has a bitter taste but delicious in meals bitter leaf in Nigeria and Africa generally) on the due to its pleasant nostalgic bitterness when it interacts antibacterial activity of the plant. Use of plant extracts or their active principle may ethanol extracts of these plant leaves were tested against serve as source of new drugs or sources of intermediate Escherichia coli. 100 g of the leaf (in three batches) were anti-helmitic and anti-malaria properties (Abosi and pounded using sterile mortar and pestle. Bitter leaf (Vernonia amygdalina) amygdalina (Bitter leaf). food or as an herbal medicine without appreciable toxicity 1995). loss of memory. crayfish. itching. dry matter of 17·02%. amygdalina fresh leaf had moisture content of aluminum foil and left for 24 h in the refrigerator. the final volume of the soup was 100 ml.

biochemical and antibiotic RESULTS susceptibility. Bayelsa The plates were inoculated at 37°C for 72 h to observe state and the Department of Pharmaceutical Microbiology the zones of inhibition. aureus and E. coli) .coli S. aureus and E. the hot water extract discs (Kigigha and Atuzie. coli were obtained agar plates to avoid overlapping of the zones of inhibition.1 filter paper and the extract stored in a borer. From Figure 1. respectively from Gloryland Clinic. The discs were well spaced in the Clinical isolates of S.1 ml each) refrigerator for use. 1 filter paper using cock Whatman No. extracts including the one percent Ampiclox control were (in triplicates) placed on separate Nutrient agar plates Test bacterial isolates which had been confluent inoculated with the test E. Effect of Bitter leaf on test organisms (S. They were characterized with respect to their morphological. aureus isolates. 2010). indicated that all the Preparation of antibacterial discs from leaf extracts three extracts were inhibitory to the two bacterial test isolates viz. College of Health Sciences. aureus (which represent the The disk-diffusion technique was used to test the enteric forms and the superficial etiologic agents antibacterial activity of the different extracts using 10 mm respectively). were impregnated on the disc in triplicates using sterile pipette and air. coli and S. (P = < 0·001). Hot W = Hot water Extract. E. BLS= Bitter leaf Soup Extract AMPI = Ampiclox Extract. The result as shown in Figure 1. coli and S. The solution containing the extracts (0. Bayelsa state. Kigigha and Onyema 43 Antimicrobial Activity of Bitterleaf 60 E. This was strained with sterilized and then filtered using prepared from Whatman No. Yenagoa.Et Ex = Ethanol Extract. Antibiotic discs were indicated the highest inhibitory zone on E. Laboratory.aureus 50 Zone of Inhibition mm 2 40 30 20 10 0 Et Ex Hot W BLS AMPI Treatment of Bitterleaf Figure 1. coli and also . Niger Delta University.dried. Media and apparatus Antibacterial assessment of extracts All media were prepared according to manufacturer’s specifications while all glassware were washed with Using flamed-sterilized and cooled pair of forceps the detergent and rinsed with deionized and distil water three discs which had been impregnated with the bitter-leaf times and were then autoclaved for 15 min. n = 3. Data = (Mean = ± SE)..

Res. aureus compared to the Ampiclox control (P < using SigmaStat statistical package. All pair wise multiple 0.025). Amoxicillin.001). AMPI Sa (P = <0.050. Et Ex Sa (P = 0.001). Hot W Sa vs. +++ = highly positive. AMPI Ec Ampiclox on E. Test and Assessment Staphylococcus aureus Escherichia coli Colony Morphology Cocci Rod Gram stain + ve . BLS Ec vs. Gentamicin and Norfloxacin. coli.001) The antibiotic resistance pattern indicated that the test . on S. Sa (P = 0. Ec (P = 0. out of the ten antibiotics tested viz.001). Et Ex Ec accordingly.001). Antibiotics sensitivity of the test bacterial isolates.001). -ve: = Negative reaction. In Table 1. BLS Sa (P <0. AMPI Sa (P = <0.004).001). Hot W Sa vs. Hot W Ec vs. Statistical analysis The differences in the mean values among the treatment Discussion groups were greater than would be expected by chance. AMPI Ec While the Staphylococcus isolate was resistant to three (P = 0. Table 1. AMPI Cephlexin and Ominoxazole. Table 2. Tetracycline. +ve: = Positive reaction.ve Growth in MSA + ve NT Coagulase test (tube/slide) + NT Catalase test + NT Sugar Utilization  Lactose NT +  Mannitol NT +  Glucose NT +  Sucrose NT + EMB NT + CLED NT + Key: NT: = Not Tested. Ampicillin. Microbiol. Clindamycin. AMPI Sa (P = <0. ++ = moderately positive. (P = 0. BLS Ec vs. Escherichia coli Staphylococcus aureus Antibiotics Sensitivity Antibiotics Sensitivity Nitrofurantion (N) ++ Ofloxecin (OF) +++ Ciprofloxacin (CIP) +++ Erythromycin (E) +++ Tetracycline (TE) R Ciprofloxacin (CIP) +++ Amoxicillin (AX) R Clindamycin (CD) R Ofloxacin (OF) ++ Gentamycin (GN) +++ Chloramphenicol (C) + Cephaplexin (CX) R Cefuroxine (CF) R Ominoxazola (CE) R Ampicillin (AM) R Ampicillin/ CLOXAC ++ Gentamicin (GN) R Floxapen (FX) + Norfloxacin (NB) R Augumentin (AU) +++ Key: + = Slightly positive.001) and also higher than the inhibitory effect of Hot W Ec vs. the test isolates were characterized Hot W Ec vs. (P<0. Morphological and biochemical characteristics of the test bacterial isolates. R = Resistant. Hot W Ec vs. indicated that E. AMPI Sa (P <0. Hot W Ec vs.025).036). there was statistically significant difference (P < 0. aureus (P < 0.44 Sky.001). The antibiotic resistance pattern in Table 2. Et Ex Sa vs.036). Hot W antibiotics tested viz. This was followed by the inhibitory effect of bitter comparison procedures at level of Comparison P < 0. leaf soup extract on E.001). AMPI Sa (P = <0. which was significantly higher (or Tukey Test) indicated as follows: than the inhibitory effect of Ampiclox on S. coli was resistant to six out of the ten Hot W Ec vs.002). Cefuroxime. J. coli (P = 0. Et Ex Ec vs.

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