antibacterial effect of Azadirachta indica (neem plant) against Pseudomonas
aeruginosa, Klebsiella ozanae, Staphylococcus aureus
and Escherichia coli was determined using the agar cup plate
techniques. The phytochemical components of A. indica showed the presence of
saponin and phlobatanin and the absence of alkaloids, tannis, phenolics,
plycosides, flavonoids, and triterpenes, the result showed that the test
organisms were susceptible to 500mg/ml, 50mg/ml and 5mg/ml of the plant
extract. The minmum inhibitory concentration (MIC) and minimum bactericidal
concentration (MBC) were determined, the result showed that the MIC and MBC
were 5mg and 50mg respectively for Pseudomonas aeruginosa, K.
ozanae, Staphylococcus aureus and Escherichia coli.
The result of the study suggest that extracts of A. indica could be suitable
for the treatment of various infections caused by P. aeruginosa, K.
ozanae, S. aureus and E. coli.
of Content vi-vii
and methods 8
and preparation of samples 8
of specimen 8
of materials 8
screening of extracts of A. indica 8
susceptibility test 9
of minimum inhibitory concentration (MIC) 9
activities of the extracts 10
activities of the extracts 10-12
4.0 Discussion 13-14
5.0 Conclusion 15
Plants contain many biologically
active compounds which have potential for development as medicinal agents. Herbal
medicines already form the basis of therapeutic use in the developing countries
but on recent there has been an increase in the use of herbal medicine in the
developed world too (De and Ifeoma, 2002; El-manhood et al, 2010) plants
provide alternative strategy in search for new drug. There is a rich abundance
of plant reported in traditional medicine to process protective and therapeutic
properties (Kayode and Kayode, 2011). It is likely that plants will continue to
be a valuable sources of new molecules which may, after possible chemical manipulation, provide new and
improve drugs (Shah et al; 2006) Bacterial resistance to antibiotic
represents a serious problem for clinicians and the pharmaceutical industry and
great efforts are being made to reverse this trend and one of them is the widespread
screening of medicinal plant from the traditional system to medicine hoping to
get some newer, saver and more effective agents that can be used to fight
infectious diseases (Natarian et al, 2003)
Azandirachta indica is one of such medical plants
belonging to the meliaceae family and the indigenous to Southern Asia (Akula et
al, 2003) Azadirachta indica commonly known as neem
has attracted worldwide prominence in recent years, owing to its wide range of
medicinal properties Neem has been extensively used in Ayurveda Umani and homoeopathic
medicine and has become a cynosure of modern medicine
Neem elaborates a fast array of
biologically active compound that are chemically diverse and structurally
More than 140 compounds have
been isolated from different parts of neem. All parts of the neem tree leaves,
flowers, seeds, fruits and roots and sac bark have been used traditionally for
the treatment of inflammations infections, fever, skin diseases and dental
disorders. The medicinal utilities have been described especially for neem
leaf. Neem leaf and its constituents have been demonstrated to exhibit
antihyperglycaemic, Antiurcer, antimalaria, antifungal, antibacterial,
antiviral, antiocidant, antimutagenic and anticarcinogenic properties (Talwar et
al;1997 Biswas et al 2002; Subapriya and Nagine; 2005).
The objective of this study
therefore are to determine the phytochemical components of the leaf extract of A.
Indica to determine the minimum
inhibitory concentration (MIC) of the extract on Pseudomonas aeruginosa,
Nebsiella Ozanas, Staphylococcus aureus and Escherichia
1.1 LITERATURE REVIEW
Plants contain many biologically active compounds
which have potential for development as medicinal agents.
Herbals medicines already form the basis of therapeutic use
in the developing countries, but of recent there has been an increase in the
use of herbal medicines in the developed world too (De and Ifeoma, 2002). The
photochemical compound of the A. Indicial have been established in previous
studies and these include tannins, saponins, alkaloids, carbohydrate, phenols,
flavonoids, anthraquinones, cardiac glycosides, sterol and resins (Sundarasivara
and Nazma, 1977; Bhomick and choudhary, 1982; Rao et al, 1986; Natarajan
et al, 2003; De and Ifeoma 2002; Biswas et al, 2002). Several
studies have linked presence of these bioactive compound is plant material to antimicrobial
presence of these secondary metabolites in plants, produce some biological
activity in man and animal and it is responsible for their use as herbs. These
compounds also have served to protect the plant against infection by micro
organism predation by insect herbivores while some give plants their odours and
or flavors and some still are responsible for their pigment (Ketkar et al,
1995; El-Mahmood et al, 2008).
In some case the activity has
been associated with specific compound or classes of compounds. These active
constituents can be used to search for bioactive lead compounds that could be
used in the partial synthesis of more useful drugs (Ogbonnia et al, 2008).
In this study, a variety of pathogenic bacteria implicated
as causative agents of eye and ear infections were selected for the screening
for antibacterial activity of the crude neem seed extracts to perceive the
efficacy, anti bacterial spectrum as well as authenticate. Some of the ethnomedicinal claims the susceptibility of
the test bacteria [(A1 (Escherichia coli) B1 (P.
aeruginosa) C1(S. pyogones) and D1 (S.
aureus)] of known sensitivity extracts from both hexane and aqueous
solvents. Inhibited the growth of both the test and control bacteria though to varying
degree. In a similar study involving some dermatophites, Natatajam et al (2003)
did not record any activity for their aqueous extracts, containing to the data
present in this study, De and Ifeoma (2002) also did not record any
antibacterial activity with the aqueous extracts of both the bark and the
leaves of the neem bark extracts against their test bacteria. The zones of
growth inhibition recorded for the methanol and acetone extract by De and
Ifeoma (2002) were also smaller in size than those to influence field and
biological activities of plant base products including the age of the plant
time of harvest, drying and process of the materials method of extraction and
the solvent used.
Some antibacterial effects of
the neem seed have also been reported against S. mutans, S.
faecalis, M. tuberculosis, V. cholera,
S. pyogenes and K. pneumonia (Biswas et
al, 2002). In another study, neem seed extract have also been observe to
inhibit the growth and development of asexual and sexual stage of drug
sensitive and malaria resistance human
malaria parasite (P. falciparum), some fungi and
some viruses (Natarajan et al; (2003). This broad spectrum activity of
crude neem extracts have been linked to the presence of bioactive compounds
notably azadiractin, gedunin, nimbidin, mahmoodin and nimbolide (Biswas et
al; 2002) which makes the neem plant useful for the treatment of various
infectious conditions including those of the eye and ear, amongst the test
bacteria, S. aureus (D1) was the most susceptible, closely
followed by S. pyogenes (C1), while E. coli
(A1) and P. aeruginosa (A1) were less susceptible as shown
by their relatively smaller sizes in several authors have also reported that
plant extracts are more effective against grain-positive than gram negative
bacteria and attributed this to the differences in their cell wall structures
(Rabe and Van Staden 1997, Parekh and Chanda, 2006), the control bacteria were
more susceptible to the toxic effects of the crude extracts than the test
bacteria through the sensitivity also varied according to strains.
The effects of PH on
the efficacy of the crude extracts are seen. The activity of the crude seed
extracts are seen. The activity of the crude seed extracts of A. indica was
optimal under acidic PH (PH2).
The effectiveness of the crude
extracts was observed to decrease as the PH was raised to alkalinity
(PH 10). This pattern is similar for all the other bacteria and also
when hexane was used as a solvent. Activity at acidic PH is
indication of acid stability while diminished activity at alkaline PH
indicates less efficacy under alkaline conditions.
Practitioners of traditional medicine usually add some when
treating their crude extracts. Acid and alkaline treatment was carried out in
order to stimulate the situations in the stomach and the gastrointestinal
tract, because the crude seed extracts are also taken orally, in addition to
their being crashed and the juicy contents squeezed at the infections sites.
The effect of temperature on the efficacy of the crude seed
extracts on the test bacteria as seen indicated that activity were more under
elevated temperatures and this trend is similar for all bacteria, regardless
whether water or hexane was used as a solvent. This supports practices of
traditional healers who most of times boils the plant extracts to high
temperature could inactivate volatile compounds and free radicals (Marjumdar et
al; 1998). The traditional medicine practitioners were reported to
sometimes crash the seeds before squeezing the fresh juices in to the affected
eye and ear and after several applications, may achieve the antibacterial
dosage at the infections sites; this repeated application at an infectious site
is due to the lower activity of the extracts at lower temperature as depicted.
The standard antibiotic chloranphenicol, demonstrated
highest activity then the crude extracts as shown.
This is because the antibiotic is in pure state and has
undergone some refining processes that have established it as standard
antibiotic (Prescott et al; 2005), the observed differences in efficacy
may also be due the fact the extracts were in a crude form and would contain
some inert substances which do not have any antibacterial activity.
The organisms used for the purposes of controls were
consistently more susceptible than the test organisms. The quantitative measure
of the in-vitro activity of antibiotics and non-antibiotic antibacterial agents
including those agents of plant origin with anti bacterial potentials are the
MIC and MBC as shown.
The growths of the organisms were
inhibited at concentrations that ranged between 3.17-25 mg/ml for aqueous
extract and 1.59-12.5 mg/ml for hexane extract. The study showed that E.
coli (A1) and P. aeruginosa (B1) had higher MIC
values, meaning that higher concentration of the extracts are required to
inhibit the growth of these bacteria, while S. pyrogenes
(C1) and S. aureus (D1) had lower MIC values and would
require low extracts concentration to inhibit their growth and these corroborated with data presented in
plant based antimicrobial substances generally have higher MIC and MBC values
when compared to antimicrobial substances obtained from micro organism or those
that are synthetically produced because of the presence of impurities, the MBC
of the plant seed extract ranged between 6.25-50 mg/ml for hexane extracts. In this study P.
aeruginosa (B1) and E. coli (A1) had higher MBC values, thus
suggesting lower susceptibility to the crude extracts and lower MBC values for S.
aureus (D1) S. pyrogenes (C1) thus suggesting
higher activity of the extracts.
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