Antibacterial Activity of Moringaoleifera Seed Extracts ...

Antibacterial Activity of Moringaoleifera Seed Extracts On Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus Bello, S.A1* and Jamiu, A.T1 [email protected] 1 Department of Biological Sciences, Al-Hikmah University, P.M.B. 1601 Ilorin, Kwara State, Nigeria.

Abstract: This study was designed to evaluate the antibacterial activity of cold, hot aqueous, ethanolic and methanolic extracts of Moringa oleifera seeds on three bacteria of clinical significance: Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The proximate analysis of the dried seed revealed the presence of moisture (04.375±0.03%), ash (03.620±0.06%), fat (27.365±0.3%), crude fibre (20.944±0.2%), carbohydrate (05.471±0.6%) and high protein content of 49.168±0.6%. The phytochemical investigation of the cold aqueous extract revealed the presence of saponins, tannins and glycosides but the absence of flavonoids and alkaloids. The cold aqueous extract had no effect on Staphylococcus aureus but inhibited the growth of Escherichia coli and Pseudomonas aeruginosa at all concentrations. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were found to be highly susceptible to the methanolic extracts at 200mg/ml with inhibitory zones of 20.0mm, 18.0mm and 15.0mm while the highest inhibitory zone of 23.0mm was recorded for Amoxycillin (Control). The methanolic extract was bacteriostatic at 50mg/ml on Escherichia coli and Staphylococcus aureus while reaction was recorded for the cold aqueous extract. The hot aqueous, ethanolic and methanolic extracts were bactericidal at 200mg/ml except for the methanolic extract which was bactericidal at 100mg/ml on Escherichia coli. This study showed that the hot aqueous, ethanolic and methanolic extracts of Moringa oleifera seeds possess antibacterial activity on the selected bacteria and thus confirm the efficacy of Moringa oleifera seed in the treatment of infections caused by these bacteria. Keywords: Antibacterial activity,Escherichia aeruginosaand Staphylococcus aureus.

coli,extracts,inhibition,Moringa

Introduction he need for new antimicrobial agents is closely linked with the problem of emergence of strains that are resistant to most synthetic antibiotics. This has arisen due to extensive use of antibiotics, which renders most of the current antimicrobial agents inefficient in controlling some bacterial diseases (Gustavo et al.,2010). There is increased evidence to proof that medicinal plants may represent an alternative treatment for non-severe cases of infectious diseases. They could also serve as possible source of new and cheap antibiotics to which pathogenic strains are not resistant. Several works have provided the scientific basis for the popular use of plants against infectious diseases (Kitula, 2007).According to World Health Organization medicinal plants would be the best source to obtain a variety of drugs. In developed countries about 80% of plants are used in traditional medicine (Adamu et al., 2005). Therefore, such plants have been investigated for better understanding of their medicinal properties. The antimicrobial properties of many plants have been investigated by a number of researcher worldwide (Adamu et al., 2005).

T

*Corresponding author:

[email protected]; Bello, S.A1* Copyright © 2017 Nigerian Society for Microbiology

oleifera,

Pseudomonas

Herbal medicine can be used as an alternative to some commercial drugs. Medicinal plants provide immeasurable projections for new drug discoveries because of the matchless availability of chemical range (Sasidharan et al., 2011). Medical uses of plants range from the administration of the roots, barks, stems, leaves and seeds to the use of extracts and decoction from the plants (Ogbulie et al., 2007). Plants have the ability to synthesize aromatic substances such as phenolic, (phenolic acids, flavonoids, quinines, coumarins, lignans, stilbenes, tannins), nitrogen compounds (alkaloids, amines), vitamins, terpenoids (including carotenoids) and some other endogenous metabolites. These substances serve as plant defense mechanisms against predation by microbes, insects, herbivores (Bharathi et al., 2011). There is a need to focus on the alternative sources of the antibiotics as pathogenic microbes are gaining resistance against standard antibiotics (Tarun et al., 2012). Medicinal plants such as Moringa oleifera, Ocimumgratissimumand Anacardiumoccidentale have been ascertained to provide various culinary and medicinal properties. These medicinal properties exert bacteriostatic and bactericidal effects on some bacteria (Okeke et al., 2008). Moringa oleiferais an aboriginal plant of Indian subcontinent and has become naturalized in the tropical and subtropical areas around the world. Nearly thirteen species of Moringa oleiferaare included in the family Moringaceae (Fozia et al., 2012).Moringa oleifera seeds are considered to be antipyretic, acrid, bitter (Oliveira et

Nigerian Journal of Microbiology 2017, 31(1): 3873-3881 Published online at www.nsmjournal.org

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al., 1999) and reported to have some antimicrobial activity. Moringa oleiferapossessess antibacterial and antifungal, anti-helmintic, antiviral, antifibrotic, antiinflammatory, anti-hyperglycemic, anti-tumour, anticancer, anti-clastogenic, analgesic, antipyretic, antihypertensive and anti-fertility properties (Fozia et al., 2012).

Test for flavonoids One (1.0) mls of aqueous extract was added to one (1) ml of 10% lead acetate solution. The formation of a yellow precipitate was an indication for the presence of flavonoids (Sofowora, 1993).

Material And Methods Collection and Maintenance of Test Organisms Already characterized and identified pure clinical isolates ofStaphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were obtained from the Department of Medical Microbiology and Parasitology, University College Hospital, Ibadan, Oyo state. The isolates were maintained on nutrient agar slants at 4oC until required for use. Plant sample collection The seeds of Moringa oleiferawere collected along University of Ilorin road, Tanke, Ilorin, Kwara state. The sample was transported to the laboratory for preparation and analysis. Sample Preparation and Extraction The seeds were dried under shade for 7 days and pulverized using a clean sterile electric blender into powdered form. About forty gram (40g) of the powdered seed was soaked in three hundred (300ml) of each solvents namely methanol, ethanol, cold and hot water. Each conical flask was covered with cotton wool wrapped with aluminium foil and was left in a rotary shaker for two days. The extract was filtered using sterile filter paper (Whattman No 1). The filtrate was evaporated to dryness using a water bath at 70oC for two days. The extract was scrapped into a sterile sample bottle using a sterile spatula (Mashiar et al., 2009). Proximate analysis of crushed seeds The proximate analysis was carried out at the chemistry laboratory, Nigeria Stored Product Research Institute (NSPRI), Ilorin, Kwara State. The proximate composition of the crushed seeds were determined by the method described by the Association of the Analytical Chemist (AOAC, 2005). The proximate analysis that were carried out were moisture, crude fat, ash, protein and carbohydrate content determination (Orhevba et al., 2013). Phytochemical Screening of the Extracts Test for tannins Two (2.0) mls of aqueous extract was stirred with two (2.0) ml of distilled water and few drops of FeCl3 solution were added. The formation of a green precipitate was an indication for the presence of tannins (Sofowora, 1993). Test for saponins Five (5.0) mls of aqueous extract was shaken vigorously with five (5.0) ml of distilled water in a test tube and warmed. The formation of stable foam was an indication for the presence of saponins (Sofowora, 1993).

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Test for alkaloids Three (3.0) mls of aqueous extract was stirred with three (3) ml of 1% HCl on a steam bath. Mayer’s and Wagner’s reagents were added to the mixture. Turbidity of the resulting precipitate was an indication for the presence of alkaloids (Sofowora, 1993). Test for cardiac glycosides Two (2.0) millilitres of each extract was dissolved in two (2) mls of chloroform; two (2) mls of sulphuric acid was added carefully and shaken gently. A reddish brown colouration was an indication for the presence of a steroidal ring (Sofowora, 1993). Reconstitution of Extracts Two (2) g of each of the four extracts was dissolved separately in l0mls of distilled water to give concentration of 200mg/ml,lower concentrations were obtained by diluting the extracts to obtain concentrations of l00mg/ml, 50mg/ml and 25mg/ml. The tubes containing the various concentrations were labelled and used immediately (Banso and Ayodele, 2001). Preparation of Sensitivity discs with extracts Discs of 6mm in diameter were punched out using a cork borer and placed in a sterile glass petridish, the discs were sterilized in an oven at 170oC for about 30 minutes after which they were allowed to cool.The already sterilized paper discs were soaked in the various concentrations of the extract. Twelve (12) discs were soaked into each concentration. The discs were allowed to absorb the solution and were kept for further analysis (Bauer et al., 1966). Antibacterial assay using Disc diffusion method This was done using the disc diffusion method as described by Kirby-Bauer, 1966. Mueller Hinton agar medium was poured into sterile petri dishes and allowed to solidify. Ten (10) mlsof 0.5 McFarland standards of test organisms was poured on the solidified agar and was spread all over the surface of the agar. The soaked discs were picked using sterile forceps and were dropped on the surface of the agar. The plates were incubated for 24 hours at 37oC. The zone of clearance was measured using a meter rule. Controls were set up with discs soaked with different solvents namely; hot and cold water, ethanol and methanol with no plant extract (Bauer et al., 1966). Determination of MIC of the Extracts The minimum inhibitory concentration of the plant extracts were determined using Broth dilution method (Santosh, 2013). One (1.0) ml of the diluted extracts concentration of 200mg/ml, 100mg/ml. 50mg/ml and 25mg/ml was mixed with 9ml of Mueller Hinton broth in different test tubes. The contents were thoroughly

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mixed and the tubes were inoculated with 0.1ml of 0.5 McFarlandstandards of the test isolates. The tubes were incubated at 37oC and examined for turbidity after 24 hours. The least concentration of the extract that did not permit show turbidity of the inoculated test isolates in the broth medium was regarded as the MIC in each case. Test tubes inoculated with the test isolates without the extracts served as controls (Santosh, 2013).

presence of moisture, protein, ash, fat, crude fibre and carbohydrate in the seed. The result indicated that the dried Moringa oleifera seeds contain appreciable amount of protein (49.168±0.6%). Table 2 shows the phytochemical analysis of the cold aqueous extract of Moringa oleifera seed. The presence of saponins, tannins and cardiac glycosides were observed. Amongstthe observed phytochemicals, flavonoids and alkaloids were absent. Table 3 shows the antibacterial activity of cold aqueous, hot aqueous, ethanolic and methanolic extracts of Moringa oleifera seed on test organisms. For the cold aqueous extracts the results showed that at concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml, Escherichia coli was susceptible with zones of inhibition of 11mm, 11mm, 10mm and 10mm respectively. Similarly, Pseudomonas aeruginosawas susceptible with inhibitory zones of 11mm, 10mm, 10mm and 9mm respectively. Staphylococcus aureuswas not affected by all concentrations of the cold aqueous extract. For the hot aqueous extracts the results showed that at concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml, Escherichia coli was susceptible with zones of inhibition of 12mm, 11mm, 10mm and 9mm respectively. Similarly,Pseudomonas aeruginosawas susceptible with inhibitory zones of 11mm, 10mm, 10mm and 9mm respectively. Staphylococcus aureuswas susceptible to all concentrations of the hot aqueous extract with inhibitory zones of 11mm, 11mm, 11mm and 10mm respectively.

Determination of MBC of the Extracts The concentrations of the extract that showed no visible growth after incubation were inoculated on Mueller-Hinton agar plates and incubated at 37oC for 24 hours. The lowest concentration of the extract that showed no growth on the plate after 24 hours were taken as the minimum bactericidal concentration (Auwal et al., 2013). Antibiotic Susceptibility Test This was done as described by National Committee for Clinical Standards (NCCLS) (2002) using Agar disc diffusion method. Dried surface of MuellerHinton agar plate was swabbed with 0.5ml broth culture of the standardized test organisms. Commercially purchased antimicrobial discs were placed onto the surface of the inoculated agar plate and the plates were incubated at 37oC for 24hours. Results Table 1 shows the proximate analysis of the crushed Moringa oleiferaseeds. The results revealed the Table 1. Proximate analysis of crushed seed TEST

OBSERVATION

Saponins

Stable foam formed

+

Green precipitate formed Absence of deep orange colour

+ _

Tannins Flavonoids

INFERENCES

Alkaloids

A clear solution formed

_

Cardiac glycosides

Steroidal ring formed

+

Table 2. Phytochemical screening of aqueous extract ofMoringa oleiferaseedParameter Moisture

Composition (%)

Composition (%)

Mean ± S.E.M (%)

04.348

04.401

04.375±0.03

Protein

49.786

48.549

49.168±0.6

Ash Fat Crude Fibre

03.676 27.083 20.784

03.563 27.647 21.104

03.620±0.06 27.365±0.3 20.944±0.2

Carbohydrate

05.677


05.264

05.471±0.2

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Table 3. Antibacterial activity of cold aqueous, hot aqueous, ethanolic and methanolic extracts of seeds of Moringa oleifera on test Test organisms

Zone of inhibition (mm)

MSC (mg/ml)

MSH (mg/ml)

MSE (mg/ml)

MSM (mg/ml)

200

100

50

25

200

100

50

25

200

100

50

25

200

100

50

25

Escherichia coli

11

11

10

10

12

11

10

09

13

11

11

10

18

16

15

12

Pseudomonas aeruginosa

11

10

10

9

11

10

10

09

13

12

11

11

20

15

14

12

Staphylococcus aureus

0

0

0

0

11

11

11

10

11

10

10

09

15

12

12

11

Keys: MSC= Moringa oleiferaSeed Cold aqueous extracts MSH= Moringa oleiferaSeed Hot aqueous extracts MSE= Moringa oleiferaSeed Ethanolic extracts MSM= Moringa oleiferaSeed Methanolic extracts For ethanolic extracts the results showed that at concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml, Escherichia coli was susceptible with the zones of inhibition of 13mm, 11mm, 11mm and 10mm respectively. Similarly,Pseudomonas aeruginosawas susceptible with inhibitory zones of 11mm, 10mm, 10mm and 9mm respectively. Staphylococcus aureuswas susceptible to all concentrations of the ethanolic extract with inhibitory zones of 11mm, 10mm, 10mm and 9mm respectively. For the methanolic extracts the results showed that at concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml, Escherichia coli was susceptible with zones of inhibition of 18mm, 16mm, 15mm and 12mm respectively. Similarly,Pseudomonas aeruginosawas susceptible with inhibitory zones of 20mm, 15mm, 14mm and 12mm respectively. Staphylococcus aureuswas susceptible to all Published by Nigerian Society for Microbiology

concentrations of the methanolic extract with inhibitory zones of 15mm, 12mm, 12mm and 11mm respectively. Comparison between the antibacterial potential of the extracts and standard antibiotics on test organisms revealed that methanolic extracts had the high zone of inhibition (20mm) at 200mg/mlonPseudomonas aeruginosa while highest inhibitory zone of 23.0mm was recorded for Amoxycillin (Control) against Pseudomonas aruginosa.

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Table 4 shows the minimum inhibitory concentration of Moringa oleifera seed extracts on test organisms. There was no minimum inhibitory concentration for the cold aqueous extract on the three test organisms while a minimum inhibitory concentration of 100mg/ml was recorded for the hot aqueous extract. The minimum inhibitory concentration of the ethanolic and methanolic extracts was 100mg/ml except for Escherichia coli and Staphylococcus aureus with minimum inhibitory concentration of 50mg/ml. Table 5 shows the minimum bactericidal concentration of Moringa oleiferaseed extracts on test organisms. Since there was no minimum inhibitory concentration of the cold aqueous extract on the test organisms, there was also no minimum bactericidal concentration of the cold aqueous extract on test organisms. The minimum bactericidal concentration of the hot aqueous, ethanolic and methanolic extracts on the test organisms was 200mg/ml except for the methanolic extract with minimum bactericidal concentration of 100mg/ml on Escherichia coli.

flavonoids and alkaloids as shown in Table 2. Vinoth et al. (2012) also reported the presence of tannins, they are a group of polymeric phenolic compounds that are able to inactivate and kill microorganisms and are used in the treatment of varicose ulcers, haemorrhoids, minor burns, as well as inflammation of gums. Herbs that have tannins as their component are stringent in nature and are used for treating intestinal disorder such as diarrhea and dysentery. The presence of saponins is in contrast to the study conducted by Jennifer and Anchana (2014) but correlate with the findings of Vinoth et al. (2012). The presence of saponins in plants has been reported to be responsible for the tonic and stimulating activities observed in Chinese and Japanese medicinal herbs. Galeotti et al. (2008) reported that saponins possess antioxidant, anti-inflammatory, antiapoptosis and immunostimulatory properties. The presence of glycosides correlates with the study done by Jennifer and Anchana (2014) and the absence of alkaloids and flavonoids also conform to the findings of Bhumika and Bijal (2015).The results obtained in this study suggest that the identified phytochemical compounds may be the bioactive constituents responsible for the efficacy of the Moringa oleifera seeds. The presence of some of these constituents have been confirmed to have antimicrobial activity (Odebiyi and Sofowora, 1978), hence it could be inferred that the seed extracts could serve as alternativeto standard antibiotics (Kubmarawa et al., 2007). The cold aqueous extract had no effect on Staphylococcus aureus at different concentrations but inhibited the growth of Escherichia coli and Pseudomonas aeruginosa. The non-activity of the cold aqueous extract against Staphylococcus aureus is in agreement with previous works which showed that aqueous extracts of plants generally exhibited little or no antimicrobial activities (Ashafa et al., 2008), but is in contrast to the findings of Ijeh et al. (2005) who reported that the aqueous extracts were more effective than the ethanolic extract.Furthermore, most researchers (Paz et al. (1995), Vlientinck et al. (1995), Martin and Eloff (1998) have generally reported that water extracts of plants do not have much activity against bacteria. The reason might be that water extracts which is different from other solvents do have myriads of compounds that may interact antagonistically in their overall activities (Busani et al., 2011). It is also suggested that the active constituents from plant materials are not readily extractable in water. Although in this study the hot aqueous extract possessed antibacterial activity on the three test organisms, methanol was a better solvent in extracting the active constituents from the seeds of Moringa oleifera.The ethanolic extracts of Moringa oleifera seed inhibited the growth of Escherichia coli, Pseudomonas aeruginosaand Staphylococcus aureus at all concentrations with inhibitory zones of 12.0mm, 11.0mm, and 11.0mmrespectively at 200mg/ml. This

Discussion The antibacterial activities of cold aqueous, hot aqueous, ethanolic and methanolic extracts of Moringa oleifera seeds on Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were assayed. These clinical isolates were selected because of their clinical importance, Staphylococcus aureus is usually implicated in wound infections, Pseudomonas aeruginosa in urinary tract infections and Escherichia coli in diarrhea and other enteric diseases. The proximate analysis of the dried pulverized seed revealed the presence of moisture (04.375±0.03%), protein (49.168±0.6%), ash (03.620±0.06%), fat (27.365±0.3%), crude fibre (20.944±0.2%) and carbohydrate (05.471±0.6%). The low moisture content of the dried seed is in concordance with the value reported by Abdullahi (2000), this low moisture is essential for better storage potential since high moisture is associated with increase of microbial activities during storage (Abdullahi, 2000).The high protein content of 49.168±0.6% disagrees with the findings of Hassan and Umar (2004), the appreciable amount of protein make it a good source of supplementary protein for man. The value of the fat content reported by Orhevba et al. (2013) is in contrast to the fat content value in this study. The carbohydrate content of 05.471±0.6% is a little bit higher than 3.93% reported by Madubuike et al. (2015), this carbohydrate content is high enough to serve as good source of energy for man. Variation in proximate results may be due to differences in variety of plants, cultivation climate and ripening stage (Taylor and van Staden, 2001). The phytochemical investigation of the cold aqueous extract revealed the presence of saponins, tannins and cardiac glycosides but the absence of Published by Nigerian Society for Microbiology

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result is in concordance to that reported by Bukar et al. (2010).Pseudomonas aeruginosa and Escherichia coliwere found to have the highest susceptibility to the methanolic extracts at all concentrations with zone of inhibitions of 20.0mm and 18.0mm at 200mg/ml. Staphylococcus aureus was also found to be susceptible with inhibitory zone of 15.0mm at 200mg/ml. This result is in agreement with the findings conducted by Lar et al. (2011) who reported that the methanolic extracts are more effective compared to the ethanolic and aqueous extracts. The antimicrobial nature of Moringa oleifera seeds is attributed to the oil contained in it, which on consumption forms a thin film over the intestinal wall, thus reducing or preventing pathogens from penetrating the intestinal walls (Nwosu and Okafor, 1995). Some studies have also shown that the antibacterial activity of Moringa oleifera seeds is linked with a gum produced in the seed (Harristoy et al., 2005). Other studies have shown that the antibacterial activity is linked with the presence of various phytochemical constituents such as saponins, tannins, flavonoids, alkaloids, cardiac glycosides and terpenoids in the seed (Nepolean et al., 2009). The mechanisms of actions of these constituents have been proven via cell membranes perturbations (Esimone et al., 2006). This coupled with the action of β-lactams on the trans-peptidation of the cell wall could lead to an enhanced antimicrobial effect of the combinations (Esimone et al., 2006).According to Dahot (1998), Moringa oleiferaextracts contain small peptides which could play an important role in the plant’s antimicrobial defence system. The proteins or peptides are believed to be involved in defense mechanism against microorganisms by inhibiting the growth of microorganisms through diverse molecular modes, such as binding to chitin or increasing the permeability of the fungal membranes or cell wall (Chuang et al., 2007). This induces membrane destabilization, and bacteria are thought to be killed by the leakage of cytoplasmic contents, loss of membrane potential, change of membrane permeability, lipid distribution, the entry of the peptide and blocking of anionic cell components or the triggering of autolytic enzymes (Zasloff, 2002). Another strategy used by plants to prevent invaders is based on the localized production of antimicrobial; low molecular weight secondary metabolites known as phytoalexins (Dahot, 1998). The minimum inhibitory concentration results in this study revealed that the cold aqueous extract did not inhibit the test organisms at different concentrations. The minimum inhibitory concentration results also showed that the methanolic extracts have better inhibitory activity compared with the hot aqueous and ethanolic extracts which could be due to the fact that most of the active constituents have been extracted from the seeds (Eloff, 1998).The hot aqueous, ethanolic and methanolic extracts were all bactericidal on the three test organisms at 200mg/ml

except for the methanolic extract which was bactericidal at 100mg/ml on Escherichia coli. All the extracts except the cold aqueous extract had bactericidal activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa which are mostly known to be multidrug resistant. More interesting is the ability of the methanolic extracts to kill Escherichia coli at 100mg/ml, the ability of the methanolic extracts to kill Staphylococcus aureus and Pseudomonas aeruginosa is noteworthy even though it was at the highest concentration of 200mg/ml.The antibiotic susceptibility pattern of the test organisms were investigated using standard antibiotic discs for Gram positive and Gram negative bacteria (Table 6 and 7). Escherichia coli was reported to be most susceptible to cefuroxime and ofloxacin, intermediate tociprofloxacin but was resistant to ceftazidime,gentamicin, amoxicillinand ampicillin whilePseudomonas aeruginosa was reportedto be most susceptible to amoxicillin but was resistant to ceftazidime, cefuroxime, nitrofurantoin and ampicillin.Staphylococcus aureus was most susceptible to ofloxacin but was resistant to ceftazidime, cefuroxime, ceftriaxone, erythromycin and cloxicillin. It was observed in this study that both Gram positive and Gram negative bacteria were susceptible to ofloxacin but were resistant to ceftazidime. Another observation worthy of note is that Staphylococcus aureus was the most resistant to the antibiotics. High levels of resistance of Staphylococcus aureusto these antibiotics may be due to the production of β-lactamase which causes the hydrolysis of β-lactam ring resulting in inactivation of β-lactam antibiotics (Fuda et al., 2005).


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Table 4. MIC of cold, hot aqueous, ethanolic and methanolic extracts on test organisms Test organisms MIC value (mg/ml) MSC MSH MSE Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus

Nil Nil Nil`

100 100 100

Key: MIC= minimum inhibitory concentration extracts

MSM

100 100 100

50 100 50

Nil: No MIC value, MSC- Moringa oleiferaseed cold aqueous

Table 5. MBC of hot aqueous, ethanolic and methanolic extracts on test organisms Test organisms MBC value (mg/ml) MSH MSE MSM Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus

200 200 200

200 200 200

100 200 200

Keys: MBC- Minimum bactericidal concentration, MSH- Moringa oleiferaseed hot aqueous extracts, MSE- Moringa oleiferaseed ethanolic extracts, MSM- Moringa oleiferaseed methanolic extracts Table 6. Antibiotics susceptibility patterns of Escherichia coli and Pseudomonas aeruginosa to standard Gram-negative antibiotics. Test organisms CAZ

CRX

Zone of inhibition in mm (Interpretation) GEN CPR OFL AUG

NIT

AMP

Escherichia coli

11(R)

20(S)

11(R)

20(I)

20(S)

9(R)

16(I)

12(R)

Pseudomonas aeruginosa

0(R)

0(R)

16(S)

18(I)

16(S)

23(S)

0(R)

0(R)

Table 7. Antibiotics susceptibility of the test organisms to standard antibiotics Staphylococcus aureus Test organism Staphylococcus aureus

CAZ

CRX

GEN

0(R)

0(R)

15(S)

Zone of inhibition (mm) CTR OFL 11(R)

19(S)

AUG

ERY

CXC

10(R)

0(R)

0(R)

Keys: CAZ-Ceftazidime(30µg),GEN=Gentamicin (10µg), CPR=Ciprofloxacin (5µg), OFL=Ofloxacin (5µg), AUG=Amoxycillin/Clavulinate (30µg), NIT=Nitrofurantoin (300µg),AMP= Ampicillin (10µg), CRX=Cefuroxime (30µg), CTR= Ceftriaxone (30µg), ERY= Erythromycin (30µg), CXC=Cloxicillin (5µg), R= Resistant, S=Susceptible and I=Intermediate. NB: The antibiotics susceptibility result was interpreted according to National Committee for Clinical Standards (NCCLS) (2002).

Conclusion This study showed that Moringa oleiferaseeds extractshave antibacterial activity on Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureusindicating a potent source of new antibiotic alternative. This inhibitory action of the extracts could be attributed to the presence of phytochemical constituents in the extracts. Further research to confirm and reinforce this include subjecting the extracts to in vivo tests using experimental animals and gas chromatography mass spectrometry to detect the specific component responsible for this antibacterial property.


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