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UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668

Received: 17th Sept., 2016 Accepted: 18th Oct., 2016

Antibacterial Activities and Phytochemical Screening of Extracts of Flower of


Azadiracthta indica (Neem) against Some Selected Clinical Isolates
*1Abdullahi, A., 1Nafiu, S.A; 2Ishaq, S.A; 1Alkali, Z. D. and 3Lambu, Z. N.
1
* Department of Science Laboratory Technology, School of Technology, Kano state Polytechnic.
3
Department of Microbiology, Kano State University of Science and Technology Wudil,
2
Department of Biology, School of Sciences, FCE (T) Bichi, Kano State
Corresponding author’s address: nafiune.sn@gmail.com; 07030918094
Abstract
A study on the phytochemistry and antibacterial effects of petroleum ether, chloroform and
methanol extract of flower of Azadirachta indica on five clinical isolates viz: Proteus mirabilis,
Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella pneumoniae
was carried out using deep well agar diffusion and broth dilution methods. The results obtained
showed that methanol flower extract possessed the highest antibacterial activity against P.
miravilis with inhibition zone of 23mm at the concentration of 4000μg/ml. The minimum
inhibitory concentration (MIC) of the petroleum ether flower extract was 62.5μg/ml for all the
test organisms while P. miravilis, S. aureus and K. pneumoniae had MIC value 62.5μg/ml with
the exception of E. coli and P. aeruginosa.Methanol flower extract recorded 62.5μg/ml
against P. miravilis, S. aureus. While E. coli, P. aeruginosa and K. pneumoniae had no MIC
value across all the concentrations. the minimum bactericidal concentration (MBC) for the
petroleum ether extracts was 125μg/ml, for chloroform extracts was 250μg/ml and for methanol
extracts was 62.5μg/ml. Phytochemical analysis of the flower extracts showed the presence of
reducing sugars, alkaloids, tannins, flavonoids, resins, and saponins, in all the extracts.
Petroleum ether flower extract (PFE) showed antibacterial activities against all the test
organisms while Methanol flower extract (MFE) had antibacterial activities against only two test
organisms.
Key words: Phytochemistry, Azadirachta indica, extracts, Clinical isolates ,MIC, MBC

INTRODUCTION patient tolerance, relatively less expensive,


Medicinal plants have a long history of use acceptance due to long history of use and
and that is widespread in both developing being renewable in nature (Vermani and
and developed countries. According to Garg, 2002). It is known that more than 400,
reports of the World Health Organization, 000 spp. of tropical flowering plants have
80% of the world’s population relies mainly medicinal properties and this has made
on traditional therapies which involve the traditional medicine cheaper than modern
use of plant extracts or their active medicine (Odugbemi, 2006). Some plant
substances (WHO, 1993). Microorganisms decoctions are of great value in the treatment
have developed resistance against many of diarrhoea or gastrointestinal disorder,
antibiotics due to the indiscriminate use of urinary tract infections, skin infections,
antimicrobial drugs (Ahmad et al., 1998). infertility, wound and cutaneous abscesses
Furthermore, antibiotics are sometimes (Ergene et al., 2006). The tree, Azadirachta
associated with side effects (Cunha, 2001), indica of the family Maliaceae; popularly
whereas there are some advantages of using known as neem tree or darbejiya (Hausa) is
antimicrobial compounds of medicinal an evergreen tree, native to the Southeast
plants, such as fewer side effects, better Asia and found in most tropical countries.

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UMYU Journal of Microbiology Research
UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668
It has been in use since ancient times, to MATERIALS AND METHODS
treat a number of human ailments and also Collection of plant materials
as household pesticide (Chattopadhyayet al., The flowers of Azadirachta indica was
1993; Chattopadhyay 1996; Chattopadhyay obtained from Audu Bako Zoological and
and Bandyopadhyay, 2005). Extracts from Botanical Garden, Kano. The collected
the bark, leaves, fruits and roots have been flowers were identified and authenticated in
used to control leprosy, intestinal Herbarium, department of Plant Biology,
helminthosis and respiratory disorders Bayero University Kano. Voucher No 118
(Ketkar and Ketkar, 1995). Every part of the was given to the plant.
neem tree has been used as traditional Preparation of the plant materials
medicine for house-hold remedy against The collected sample was washed
various human ailments from antiquity. thoroughly with running tap water and
Chemical investigation on the products of finally with sterile distilled water. The
the neem tree extensively undertaken in the material was chopped into small pieces and
middle of the isolation of nimbin, the first then air dried on a sterile blotter under shade
bitter compound isolated from neem oil, for 20-30 days.
more than 135 compounds have been Extraction procedures
isolated from different parts of neem Plant extract was prepared by the maceration
(Ganguli, 2002). Neem elaborates a vast method as demonstrated by Alade and Irobi
array of biologically active compounds that (1993). 200 g of the air dried powdered form
are chemically diverse and structurally of the flower of Azadirachtaindica was
complex (Subapriya and Nagini, 2005). soaked in 250 ml of petroleum ether for
Infectious diseases are the world’s leading three days. The mixture was stirred every 24
cause of premature deaths, killing almost 50 h using a sterile glass rod. The plant
000 people every day. In recent years, drug sediment was allowed to dry and soaked in
resistance to human pathogenic bacteria has 250ml of chloroform and finally in methanol
been commonly reported from all over the solvent for three days respectively. At the
world (Piddock and Wise, 1989; Singh et al., end of extraction each extract was passed
1992; Mulligen et al., 1993; Davis, 1994; through Whatman filter paper No. 1
Robin et al., 1998). However, the situation is (Whatman, UK). The procedure was
alarming in developing as well as developed repeated three timeswith fresh volumes of
countries due to indiscriminate use of the solvents used.The alcoholic filtrates
antibiotics. The drug-resistant bacteria and obtained were concentrated in water bath at
fungal pathogens have further complicated 30°C and stored at 4°C until further use. The
the treatment of infectious diseases in dried plant extracts/fractions obtained were
immune compromised, AIDS and cancer weighed and labeled as PFE(Petroleum ether
patients (Rinaldi, 1991; Diamond, 1993). In flower extract), CFE(Chloroform flower
the present scenario of emergence of extract), and MFE(Methanol flower extract).
multiple drug resistance to human Phytochemical screening
pathogenic organisms, this has necessitated a • Test for saponins
search for new antimicrobial substances Exactly 2ml of petroleum ether extract was
from other sources including plants.Thus, vigorously shaken with distilled water and
the choice of azadirachta indicaas a plant of allowed to stand for a while. A persistent
interest for this research work was done frothing indicates the present of saponins.
based of its diverse and immense therapeutic Same procedure was repeated using
ethno medicinal values. In view of the chloroform and methanolic extracts
forgoing, this research is aimed at studying (Sofowora, 1984).
the antibacterial activity and phytochemical • Test for alkaloids
properties of extracts of flower of Half gram of petroleum ether extract was
Azadirachta indica (neem) on some selected stirred with 5ml 1% HCL on steam bath.
clinical isolates.
UMYU Journal of Microbiology Research 62
UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668
The solution was cooled and filtered.1ml of colour from rose-pink to red indicates
the filtrate was treated separately with drops anthraquinone. Same procedure was
of Mayer’s dragendoff’s and wagner’s repeated using chloroform extracts and
reagents; and formation of dirty/dark brown, methanolic extracts (Ciulie, 1984)
yellow-brown or reddish brown precipitated • Test for steroids(Salkowaski’s test)
respectively indicates the presence of Two mills of concentrated sulphuric acid
alkaloid (El-olemy et al., 1994). was added to 2ml of each extract
• Test for phlobactannins .Appearance of effervescence after which a
A 2ml of each extract was added to 5ml clear reddish brown color appeared at the
HCL.Formation of turbidity/precipitation interface confirms the presence of steroidal
indicates the presence of phlobactannins. ring (Harbone,1973).
(Sofowora, 1984). Test organisms
• Test for tannins Clinical isolates of bacteria were used for
Exactly 2ml of each extract was treated with the bioassay studies. The isolates include
3 drops of 5% ferric chloride. A dark black Proteus mirabilis, Escherichia coli,
colored precipitate in a very dark solution, Staphylococcus aureus, Pseudomonas
which gives a green-black to blue black aeruginosa,and Klebsiella pneumoniae. The
colouration on dilution indicates the isolates were obtained from microbiology
presence of tannins (Sofowora,1984). laboratory of Murtala Mohammad Specialist
• Test for reducing sugar Hospital (MMSH), Kano, Nigeria. They
One gram of each extract was weighed and were further confirmed using standard
diluted with 2ml distilled water. Fehling’s biochemical test as described by
solution(A and B) were added and the Cheesbrough (2002).The isolates were
mixture warmed. A brick- red precipitate at maintained on freshly prepared nutrient agar
the bottom of the test tube indicates reducing (oxoid) slants and kept in a refrigerator at
sugars (Brain and Turner, 1975). 4oc until required for use.
• Test for flavonoids Preparation of extract concentrations
Two grams of the petroleum ether extract This was carried out using standard method
was weighed and placed in a test tube, described by Cheesbrough, (2002). Stock
followed by the addition of 10ml of DMSO. solution of the petroleum ether crude extract,
The mixture was heated, followed by the chloroform crude extract and methanol
addition of magnesium metal and six drops crude extract were prepared by weighing
of concentrated hydrochloric acid. The 0.008g of each and dissolved in 2ml of
appearance of red colour was indicatives of dimethlysulfoxide (DMSO) in glass vial
the presence of flavonoids. Same procedure bottles.
was repeated with chloroform and methanol This gave an extract concentration of
extracts respectively (Sofowora, 1993). 8000µg/ml (stock solution).Four varied
• Test for resins: extracts concentrations (4000µg, 2000µg,
A 2.0g of each extract was dissolved in 10ml 1000µg and 500µg) were prepared from the
of ethanioc acid anhydride. One drop of stock solution (8000µg) using double serial
concentrated sulphuric acid was added. The dilution for each extract.
appearance of purple colour which rapidly Preparation of antibiotic dilution
changes to violet indicates the presence of (Standard)
resins (Evans, 1995). The antibiotic chloramphenicol was
• Test for antraquinone purchased at a registered Pharmaceutical
Two mills of petroleum ether extractwere store in Kano State, Nigeria and was
treated with 5ml of benzene. This gave two reconstituted by dissolving 0.3 mg of
layers. The clear colorless upper layer was powder in a 100 ml of distilled water so as to
pipette and the organic layer treated with get a concentration of 30μg/ml. The
3ml of 10% aqueous ammonia. Change of prepared dilution of the antibiotic was used

UMYU Journal of Microbiology Research 63


UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668
for subsequent antimicrobial test and serve Determination of the minimum inhibitory
as a positive control concentration (MIC)
Preparation of inoculums The MIC of the crude extracts was
The standardization of culture was done determined using the doubling dilution
according to the method of Baker and method of Saham and Washington
Thomsberg (1983) and CLSI in2006.Two (1990).Two milliliters of the reconstituted
mm diameter colonies of the 18 h culture of crude extract at a concentration of 500μg/ml
an organism was picked with a sterile wire was added to two milliliters of sterile
loop and immersed into a sterile bottle Mueller Hinton broth. Two milliliters of this
containing Mueller Hinton broth (Hi Media) extract concentration was transferred to
and incubated for 5 hours .Normal saline another test tube and this dilution continued
was added gradually to it so as to compare until an 8th test tube is reach, giving extract
the turbidity to that of 0.5 McFarland concentrations of 250, 62.5, 31.25, 15.6, 7.8,
standard corresponding to approximately 1.5 3.9, 1.95 and 0.98μg/ml in different test
× 108cfu / ml. This was done for each of the tubes. 0. 1 ml of an 18 h culture of bacteria
test bacteria. previously adjusted to 0.5 McFarland
Assay for antibacterial activity standard (1.5 X 108cfu/ml) was inoculated
The Boakye-Yiadom (1987) agar well into each of the test tubes and the contents
diffusion method was used to evaluate the thoroughly mixed. The tubes were incubated
antimicrobial activity of the crude extracts. at 37oC for 24 h. The 9th test tube containing
Briefly, 1.0 ml of the standard inoculum was 2 drops DMSO was served as a negative
inoculated into 90 mm sterile Petri plate, control. The 10th test tube containing a
then 19 ml sterile Mueller Hinton agar was solution of 30μg/ml of chloramphenicol
added and the plate rocked gently for 1 solution was served as positive control. The
minute for even mixing of the contents. The above procedure was followed for each of
plates were kept on a flat bench for 30 the test bacteria. The lowest concentration of
minutes to gel. The six wells were made on the extract that did not show any detectable
respective agar plate by using cork borer of growth was taken as the MIC.
4mm in diameter size. Two drops of Determination of the minimum
petroleum ether, chloroform, and methanol bactericidal concentration (MBC)
extracts at different concentration of From each of the test tubes in the MIC
4000μg/ml, 2000μg/ml, 1000μg/ml, and determination that did not show any visible
500μg/ml equivalent to a potency of 400μg, growth, 100 μl of the broth was aseptically
200μg, 100μg, and 50μg respectively were inoculated on to a sterile Mueller Hinton
introduced to their respective wells. Two agar surface and gently spread all over the
drops of 30μg/ml of chloramphenicol surfaces with a sterile bent glass rod. The
solution was served as a positive control and inoculated plates were incubated for 24hrs at
0.1 ml of DMSO as a negative control. The 37oC. After incubation, the MBC is the
plates were allowed to stand on flat bench lowest concentration of the extract that
for 30 minutes to allow diffusion into the showed no was determined at the dilution, at
agar before incubation at 37oC for 24 h. The where there was no growth (colony) on the
experiment was done for each of the extract plate (De and Ifeoma, 2002).
against each of the test bacteria and mean STATISTICAL ANALYSIS
zone diameter was recorded. Antibacterial Data obtained was subjected to statistical
activity was evaluated by measuring the analysis using one way ANOVA
diameters of zones of growth inhibition RESULTS
(Hugo and Russell, 1983; WHO, Table 1 shows the results of Preliminary
2003).These experiments were repeated for physicochemical characteristics of neem
each of the test bacteria. flower crude extracts which yielded 5.6g,
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UMYU Journal of Microbiology Research


UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668
7.8g and 14g of the petroleum ether, fractions indicates that there were variations
chloroform and methanolic extracts in the colour, odour and texture of the
respectively from the initial crude of 200g. extracts.
A physical characteristic of the extracts

Table 1: Physicochemical characteristics of the flower extracts of A. indica


Plant part solvent Initial Final Colour Odour Texture
weight (g) weight(g)
Flower Petroleum 200 5.6 Dark- Fruity Oil/gummy
ether brown
Chloroform 186 7.8 Light- Fruity Gummy
green Pleasant

Methanol 176 14 Light- Pleasant Gummy


brown fruity

Table 2, illustrates the phytochemical and Anthraquinone were absent in both.


analysis of the neem flower extracts which Flavonoids were found in PFE, MBE, and
indicates presence of reducing sugars and MFE, while Resins and Saponins were
alkaloids in all the plant extracts. Tannins present in PFE fractions only.
were present in MFE only, while Steroids

Table 2: Result of the phytochemical analysis


Secondary Extracts CFE MFE
metabolite group PFE

Reducing sugar + + +
Tannins - - +
Steroids - - -
Flavonoids + - +
Resins + - -
Saponins + - -
Anthraquinone - - -
Phlabactannins - - -
Alkaloids + + +
Key: Present (+), Absent (-)

Table 3, demonstrates the antibacterial flower extract (MFE) have antibacterial


activity patterns of different fractions of activity against onlyP. miravilisand S.
flower extracts of A. indica. The result aureus with the exception of E. coli, P
showed that petroleum ether flower extracts .aureginosa and K.pneumoniae across all
(PFE) have antibacterial activity against all concentrations. Moreover, from the result
the test organisms. Chloroform flower methanol flower extract possessed the
extract (CFE) have antibacterial activity highest antibacterial activity against P.
against all isolates with the exception of E. miravilis with inhibition zone of 23mm at
coli and P. aureginosa while methanol the concentration of 4000μg/ml.

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UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668
Table 3: Antibacterial activity of flower extracts of Azadirachta indica against the
bacterial isolates
Diameter of zone of Inhibition (mm)/Extract concentration(μg/ml)
PFE CFE MFE
Isolates 500 1000 2000 4000 500 1000 2000 4000 500 1000 2000 4000

P. miravilis 15 17 19 21 13 16 17 19 17 19 20 23
E. coli 08 10 14 15 00 00 00 00 00 00 00 00
S. aureus 14 16 17 19 12 15 16 20 13 14 15 17
P. earuginosa 07 08 09 11 00 00 00 00 00 00 00 00
K. pneumonia 08 08 09 15 08 15 20 21 00 00 00 00

Table 4, showed the activities of test organisms were susceptible to


Chloramphenicol (antibiotic) as positive chloramphenicol at 30μg/ml and resistant to
control and DMSO as negative control on DMSO.
the test organisms. It indicated that all the

Table 4: Activities of Chloramphenicol (antibiotic) as positive control and DMSO as


negative control on the test organisms.
Diameter of zone of Inhibition (mm)/ Anbiotic concentration(μg/ml)

Isolates Chloramphenicol 30μg/ml DMSO

P. miravilis 14 0
E. coli 16 0
S. aureus 19 0
P. earuginosa 14 0
K. pneumonia 9 0

Table 5, shows the Minimum Inhibitory 62.5μg/ml while E. coli and P. aeruginosa
Concentration (MIC) of different fractions were resistant. Moreover, all the isolates
of A. indica extracts against the selected tested with methanolic extracts of flower of
clinical isolates. The results showed that all A. indica also have MIC value of 62.5μg/ml
the test organisms have the same MIC of with the exception of E.coli,P. earuginosa
62.5μg/ml in the petroleum ether extract and K. pneumoniae which all of them
while P. miravilis, S. aureus and K. showed resistance to methanol flower
pneumonia tested with Chloroform flower extract.
extracts have also the same MIC of
Table 5: Minimum Inhibitory Concentration of Petroleum ether, Chloroform and Methanol
flower extracts of A. indica
Extracts concentration(μg/ml)
PFE CFE MFE
Isolates 250 125 62.5 31.3 15.6 250 125 62.5 31.3 15.6 250 125 62.5 31.3 15.6

P. mirabilis - - - + + - - - + + - - - + +
E. coli - - - + + + + + + + + + + + +
S. aureus - - - + + - - - + + - - - + +
P. aureginosa - - - + + + + + + + + + + + +
K. pneumoniae - - - + + - - - + + + + + + +
Key: + = Growth not inhibited; - Growth inhibited

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UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668
Table 6, showed the minimum bactericidal 125μg/ml while S. aureus and P.
concentration of Petroleum ether, mirabilis,have no MBC value across all
Chloroform and Methanol flower extracts concentrations of petroleum ether flower
against the test organisms. Minimum extracts(PFE).In CFE only P. mirabilis had
Bactericidal Concentration of each isolate the MBC value of 250μg/ml. However in
was determined at the lowest concentration MFE S. aureus had the MBC value of
which inhibits bacterial growth. From the 250μg/ml while P. mirabilis had the MBC
result it showed that E. coli, P. aeruginosa value of 62.5μg/ml and the remaining tests
and K. pneumoniae had the same MBC of organisms were resistant.

Table 6: Minimum Bactericidal Concentration of Petroleum ether, Chloroform and Methanol


flower crude extract of A. indica
Extracts concentration(μg/ml)
PFE CFE MFE
Isolates 250 125 62.5 250 125 62.5 250 125 62.5

P. miravilis + + + - + + - - -
E. coli - - + + + + + + +
S. aureus + + + + + + - + +
P. aeruginosa - - + + + + + + +
K .pneumoniae - - + + + + + + +

Key: + = Growth not inhibited; - Growth inhibited

Discussion pigments (Ketkar et al., 1995, El-Mahmood


In this study the phytochemical screening of et al,. 2008). In some cases, the activity has
the flower extracts of Neem plant revealed been associated with specific compounds or
the presence of reducing sugars, alkaloids, classes of compounds. These active
Tannins, Steroids, Anthraquinone, constituents can be used to search for
Flavonoids, Resins, phalabactannins and bioactive lead compounds that could be used
Saponinsin in the extracts . The in the partial synthesis of more useful drugs
phytochemical components of the A. indica (Ogbonniaet al., 2008).
have been established in previous studies A .indica flower extracts of both petroleum
and these include tannins, saponins, ether, chloroform and methanol solvents
alkaloids, carbohydrates, phenols, inhibited the growth of the test bacteria,
flavonoids, anthraquinones, cardiac though to varying degrees. Petroleum ether
glycosides, sterols and resins (Sundarasivara flower extracts showed low to moderate
and Nazma, 1977; Rao et al., 1986, activity where PFE had inhibition zone (IZ)
Natarajanet al., 2003; De and Ifeoma, 2002; between 07-21mm. Chloroform flower
Biswas et al., 2002). Several studies have extract showed low to moderates activity
linked presence of these bioactive with (IZ) between 8-21mm with the
compounds in plant materials to exception of E. coli, and P. aeruginosa
antimicrobial activity. The presence of these respectively. Methanolic extract showed
secondary metabolites in plants, produce fairly higher degree of activity in which
some biological activity in man and animals MFE had (IZ) of between 13-23mm.From
and it is responsible for their use as herbs. the result it shows that methanolic extract
These compounds also serve to protect the was the most effective with the wider zone
plant against infection by microorganisms, of inhibition of 23mm (Table.3).The
predation by insects and herbivores, while effectiveness of these solvents may be due to
some give plants their odors and or flavors their polarity where methanol was the most
and some still are responsible for their polar among the three solvents used.
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UJMR, Volume 1 Number 1 December, 2016 ISSN: 2616 - 0668

The zones of growth inhibition recorded for activity of the extracts against the
the methanol and acetone extracts by De and organisms. In this study, P. mirabilis, had
Ifeoma (2002) were also smaller in size than the lowest MBC value (125μg/ml) in
those obtained in this study. Several factors methanolic flower extracts (MFE) (Table 6).
are known to influence yield and biological The quantity of the active ingredients
activities of plant based products, including required to effect complete kill may not
the age of the plant, time of harvest, drying matter since medicinal plants have been
and processing of the materials, methods of reported to have little or no side effects
extraction and the solvents used (Hassain-Eshrat, 2002; Ogbonnia et al.,
The quantitative measure of the in-vitro 2008). The MIC and MBC values for neem
activity of antibiotics and non-antibiotic leaves against some fungal isolates were
antibacterial agents including those agents of reported to be 250μg/ml by Natarajan et al.,
plant origin with antibacterial potentials are (2003) which are in accordance with the
the MIC and MBC. The minimum inhibitory MBC values of P. mirabilis, in Petroleum
concentration was defined as the lowest ether neem flower extract and S.aureus in
concentration of the compound to inhibit the MFE obtained in this study. De and Ifeoma
growth of microorganisms .Table 5 shows (2002) reported that at a concentration of 10
the minimum inhibitory concentration (MIC) mg/ml their crude extracts were unable to
of A. indica flower extracts against the inhibit the growth of some bacteria,
selected clinical isolates and the results particularly P. aeruginosa and E. coli.
showed that all the test organisms have the However, the result of this study showed
same MIC of 62.5μg/ml in PFE, while in that PFE of A. indica inhibited the growth of
CFE P. mirabilis, S. aureus and P. aeruginosa and E. coli even at lower
K.pneumoniae had the same MIC of concentration of 125μg/ml.
62.5μg/ml. In MFE P. mirabilis, and S. The varied zone of inhibition of MBE on E.
aureus had the same MIC values of coli at concentrations of 500, 1000, 2000
62.5μg/ml . The study showed that E. coli and 4000μg in table3 disagreed with reports
and P. aeruginosa had no MIC value in CFE by Yagoub et al.,(2007) who in their
while E. coli, P. earuginosa and K. preliminary screening for anti-microbial
pneumonia also have no MIC values in MFE activity of different plants against different
at all concentrations, meaning that higher organisms, methanolic extracts of A.indica
concentration of the extracts are required to produced zero zone of inhibition against E.
inhibit the growth of these bacteria. The coli.
MBC of the PFE extracts was 125μg/ml Conclusion
against E. coli, P. aeruginosa and K. Based on the pharmacological results of the
pneumonia respectively while P. mirabilis study, it could be confirmed that the extracts
and S. aureus have no MBC across all the contain chemical constituents of
concentration. In CFE only P. mirabilis had pharmacological significance. The
MBC value of 250μg/ml while the observation that the extracts were effective
remaining test organisms have no MBC against the test bacteria suggests the use of
across all concentrations. Meanwhile in crude extract of flower of A. indica against
MFE P. mirabilis had MBC value of infection caused by clinical isolates like P.
62.5μg/ml while, S. aureus had MBC value mirabilis, Escherichia coli, S. aureus,P.
of 250μg/ml while E. coli, P. aeruginosa aeruginosa, and K. pneumonia.It is therefore
and K. pneumoniae had no MBC values at recommended for the isolation and
all the concentrations used. The higher the purification of bioactive compounds in
MBC values is the lower the susceptibility Neem tree responsible for the antimicrobial
of microorganism to the crude extracts and activity.
the lower the MBC values is the higher the

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