ABSTRACT
Aliments are parts of human life, but due to resistant nature of synthetic drugs and its cost rate there is need for alternative and there comes medicinal plants which have and continued to play a major role in treating human diseases. Synedrella nodiflora is a medicinal plant that has been used to treat constipation, dysentery and diarrhea. This study is designed to investigate the antimicrobial, antioxidant and laxative activities of synedrella nodiflora. in vitro method of assessment were used to determine the scavenging activities of the extract on 2,2-diphenyl-1-picrylhydrozyl(DDPH), Nitric oxide and Anti-lipid peroxidation as well as determination of the antibacterial activities against some bacteria strains like staphylococcus aureus, proteus mirabilis, klebsiella pneumonia, Escherichia coli, salmonella typi and pseudomonas aeruginosa. Laxative activity was also carried out on wistar albino rat in this study by diluting the ethanolic extract of the plant using serial dilution. The plant extract was diluted to different concentrations of 2000mg, 1000mg, 500mg, 250mg, 125mg, 62.5mg respectively and administrated intraperitoneally to the experimental animals. Acute toxicity study of the plant extract was determined and the LD50 was found to be (>6400). Ducox was used as control drug for laxative, vitamin c was used as control for antioxidant and ciprofloxacin was used as control for antimicrobial activities. At the end of the experiment, the result shows that the activities of the plant extract against 2,2-diphenyl-1-picrylhydrozyl(DDPH), Nitric oxide, Anti-lipid peroxidation, bacteria strains and its laxative action were all concentration dependent. The 2,2-diphenyl-1-picrylhydrozyl(DDPH), Nitric oxide and Anti-lipid peroxidation has an IC50 values of 3.96mg/ml, 4.15mg/ml and 4.38mg/ml respectively. The result of the antioxidant reveals potent free radicals scavenging activities. The laxative and antimicrobial activities show a good evidence that the crude extract of synedrella nodiflora is a good source of antimicrobial and laxative agents which justifies its traditional use.
TABLE OF CONTENT
TITLE PAGE
i
CERTIFICATION ii
DEDICATION
iii
ACKNOWLEDGEMENT
iv
TABLE OF
CONTENTS
v
LIST OF TABLES,
FIGURES AND PLATES
vii
ABSTRACT
viii
CHAPTER ONE
1.0
INTRODUCTION
1
1.1
SYNEDRELLA NODIFLORA
1
1.1.1
USES 3
1.2
JUSTIFICATION FOR THE
STUDY
6
1.3 AIMS AND OBJECTIVES
6
1.4 DEFINITIONS OF TERMS
6
CHAPTER TWO
2.0 LITERATURE REVIEW
8
2.1 THE PHYSIOLOGY AND FUNCTIONS OF
LAXATIVE 9
2.1.1
PHARMACOLOGY OF LAXATIVE
9
2.2 REVIEW OF ANTIOXIDANT ACTIVITY 10
2.3 ANTIMICROBIAL
12
CHAPTER THREE
3.0 MATERIALS AND METHODS 17
3.1 STUDY SITE
17
3.2 COLLECTION OF PLANT SAMPLE
17
3.2.1
PREPARATION OF PLANT EXTRACT
17
3.3 RAT COLLECTION AND TREATMENT
18
3.4 MEDIAN LETHAL DOSE
18
3.4.1
ACUTE TOXICITY STUDY OF ETHANOL EXTRACT OF SYNEDRELLA NODIFLORA ON RAT 19
3.5 EXPERIMENTAL INDUCTION OF LAXATIVE
19
3.6
DETERMINATION OF ANTIOXIDANT ACTIVITY
20
3.6.1 DDPH ASSAY 20
3.6.2
DETERMINATION OF INHIBITORY ACTIVITY AGAINST LIPID PEROXIDATION 21
3.6.3
IN-VITRO NITRIC OXIDE RADICAL (NO) SCAVENGING ASSAY
22
3.7 EXPERIMENTATION OF ANTIMICROBIAL
ACTIVITY
22
3.7.1
PREPARATION OF INNOCULA
22
3.7.2
ANTIMICROBIAL ACTIVITY TEST
23
3.7.3
MINIMUM INHIBITORY CONCENTRATION (MIC)
23
3.7.4
PREPARATION OF ANTIBIOTIC STOCK SOLUTION 24
3.8 IC50 CALCULATION
24
3.9 STATISTICAL ANALYSIS 24
CHAPTER
FOUR
4.0 RESULT
25
4.1 ACUTE TOXICITY AND LETHALITY TEST
RESULT
25
4.2 ANTIMICROBIAL TEST RESULTS
26
4.3 ANTIOXIDANT TEST RESULTS
29
CHAPTER FIVE
5.0 DISCUSSION AND
CONCLUSION
35
5.1 DISCUSSION
35
5.2 CONCLUSION
38
REFRENCES
LIST OF TABLES, FIGURES
AND PLATES
Figures
Figure 1: Synedrella nodiflora
Figure 2: Wistar
Albino Rat
Figure 3: Bar
chart showing the antimicrobial activity of the zone of inhibition of ethanolic
extract of Synedrella nodiflora at
2000mg and standard drug (ciprofloxacin at 500mg).
Figure 4: Bar
chart showing the mean stool weight and groups of the experimental animals in
grams.
Tables
Table 1: Result of
acute toxicity test
Table 2: Result of
minimum inhibitory concentration of the plant extract (MIC).
Table 3: Result of
antimicrobial activity of the plant extract and standard drug.
Table 4: DDPH
Scavenging activity
Table 5: Nitric
oxide scavenging activity
Table 6:
Anti-lipid peroxidation activity
Table 7: Result on
laxative activity
Plates
Plate 1: prepared
disc of different concentration of the plant extract and the control
(ciprofloxacin).
Plate 2: The zone
of inhibition of ethanolic leave extract of synedrella
nodiflora on the six test microorganism (proteus mirabilis, pseudomonas aeruginosa, salmonella typi,
staphylococcus aureus, klebsiella pneumonia and Escherichia coli)
CHAPTER
ONE
1.0 INTRODUCTION
1.1 Synedrella nodiflora
Synedrella nodiflora (L)
Gaertn. belongs to the family Asteraceae. It is a small, annual weed of
cultivation, native to tropical America, found in the plains of India and in Andaman’s.
Synedrella nodiflora is
an erect branched ephemeral herb usually 30-80 cm tall belonging to family
Asteraceae. The shallow root system is usually strongly branched. The erect or
ascending, sometimes woody stems, branch dichotomously from the base of the
plant; they tend to have long internodes and swollen nodes, to be rounded or
slightly angular in section, smooth, often finely hairy, and usually about 50
cm tall. The lower parts of the stems may root at the nodes, especially in damp
conditions. The leaves occur in opposite pairs and are 4-9 cm long, elliptic to
ovate with three prominent veins and finely toothed margins, finely hairy with
short petioles, and joined by a ridge across the stem. The flowers occur in small crowded bunches of
2-8 inflorescences at nodes and tips throughout the upper third of the plant;
each inflorescence consists of several erect bracts 3-5 mm long surrounding 5-6
marginal ray florets and 10-20 central disc florets, each 3-4 mm long with a
yellow petal. The dark brown to blackish (occasionally paler) seeds are
dimorphic. Ray floret seeds are flattened, oblong, 3-5 mm long, with
upwardly-pointing teeth along the paler marginal wing. Disc floret seeds are
thickened, elongate, 3-4 mm long, with 2-4 stiff bristles at the apex. Both
types of seed produce identical individuals, which in turn produce both types
of seed.
The seedlings have epigeal
germination. The hypocotyl is 8-19 mm long, often purplish, and hairless. The
cotyledons are elliptic, 6-8 mm long, often reddish or purplish in colour and
shortly stalked. The paired juvenile leaves are similar to the adult leaves but
smaller. It is a native to tropical
America, but now spreading pan tropically and occurring throughout the
south-east Asian region, found in the plains of India, in the Andaman’s and
West Africa. It is also found in Bangladesh, Japan, Spain, China and England.
Different activities are attributed to this vegetal species worldwide. It
contains bioactive components such as flavonoids, alkaloids, tannins etc. and
is used for the treatment of various diseases and its leaves are eaten as a
vegetable by some livestock and human.
Asteraceae family consists of
herbs, which are known to accumulate substantial amount of flavonoids and to
display anti-inflammatory, antioxidant, antimicrobial, analgesic and
antipyretic properties (Odom et al.,
2000). In Ghana, S. nodiflora (L) Gaertn. weed is used for the treatment
of epilepsy and pain (Idu et al.,
2007). In Malaysia and Indonesia, the plant is used for headaches, earaches,
stomach aches and rheumatism (Sumi et
al., 2011).
Figure1:
Synedrella
nodiflora
Scientific classification of Synedrella
nodiflora
Kingdom: Plantae
Order: Asterales
Family: Asteraceae
Tribe: Heliantheae
Genus: synedrella Gaertn
Species: S. nodiflora
Synedrella
nodiflora is a common weed of waste places and
found along the banks of rivers, streams and also along roadsides (Mshana et
al., 2000) In Ghana, the whole plant is boiled and the aqueous extract
drunk for the treatment of epilepsy whiles the leaves are used for threatened
abortion, hiccup, laxative and feed for livestock (Dalziel, 1931; Mshana et
al., 2000). The plant is also used by subsistence farmers of Ghana as
post-harvest protectants (Cobbinah et al., 1999). Traditional uses of
the plant in other African and some Asian countries have been reported. In
Nigeria, it is known that some indigenous tribes use the whole plant for the
treatment of cardiac distresses and to stop wound bleeding (Idu and Onyibe,
2007). The foliage is readily eaten by livestock in Cameroon (Irvine, 1961). In
Indonesia the young foliage is eaten as a vegetable and the leaf sap together
with other materials, is applied for stomach-ache and the plant is used in
embrocation for rheumatism (Burkill, 1985). In Malaysia, a poultice of the
leaves is used for managing sore legs and for the treatment of headache and the
sap is instilled into the ear for ear ache (Burkill, 1985). The hydro-ethanoic
extract of the whole plant has been found to possess anticonvulsant (Amoateng et
al., 2012), sedative (Woode et al., 2011), in vitro anti-oxidant
and free radical scavenging properties (Amoateng et al., 2011) and
anti-nociceptive properties (Woode et al., 2009). To explore the plant
for further pre-clinical anti-epileptic drug discovery and development, it is
important to investigate the toxicity of the plant.
1.1.1 USES
- The
young shoot of the plants can be eaten as cooked vegetable, and the shoot
are fed to pigs in Papua New Guinea.
- Crushed
leaves have been used as a treatment for rheumatism and when mixed with
those of other plants to relieve stomach pains.
- Leaves
are used as laxatives as well as in rheumatism as embrocation
- The
leaf-sap can be used for ear treatment and the oral root medicine can cure
naso-pharyngeal affections.
- The
plants can be used to cure leprosy, swellings, odema, and dropsy etc.
Medicinal plants of various types and from
different settings/habitats are of great importance to human as well as
veterinary medicine. The screening of plants extracts represents continuous
efforts to find compounds with potential to act against life-threatening
diseases such as new and/or re-emerging microbial infections (Alka and Padma,
2013). Antimicrobials of plant origin have enormous therapeutic potential and
have been used since time immemorial. They have been proved effective in the
treatment of infectious diseases and simultaneously mitigating many of the side
effects which are often associated with synthetic antibiotics (Christudas et al., 2012). On the other hand, while
it is likely that important new drugs remain hidden in plants in primary tropical
rainforest, they may also lie hidden in those belonging to other ecosystem.
There has been significant evidence that weeds
are relatively high in bioactive secondary compounds and are thus likely to
hold promise for drug discovery (Stepp, 2004). Concern has been expressed about the rising prevalence of pathogenic
microorganisms, which are resistant to the newer or modern antibiotics that have been
produced in the last three decades (Cohen,
1992; Nascimento et al., 2000).
Also, the problem posed by the high cost, adulteration and increasing toxic
side effects of these synthetic drugs
coupled with their inadequacy in diseases treatment found more especially in
the developing countries cannot be
over emphasized (Shariff, 2001).
Coincidentally, the last decade has also witnessed increasing intensive studies
on extracts and biologically active compounds isolated from plant species used
for natural therapies or herbal medicine (Nascimento et al., 2000; Rios
and Recio, 2005).
For over thousands
of years now, natural plants have been seen as a valuable source of medicinal
agents
with proven
potential of treating infectious diseases and with lesser side effects compared
to the synthetic drug agents (Iwe et al., 1999).
Laxative are foods or drugs that increases
bowel movement. Laxatives contains chemicals that help increase stool motility,
bulk and frequent, thus relieving temporary constipation. They can also be used
to ease constipations and are available over the counter (without a
prescription). (Das et al., 2010).
Certain types of foods or drugs can
promote bowel movements mainly by loosening the stool or by allowing the
constriction in the intestinal muscle to mobilize the solid waste towards the
anal opening for excretion. The primary benefits of taking a purgative
medication for providing relief from constipation (Rush et al., 2002). Laxative work in different ways and the
effectiveness of each laxative types varies from person to person which was
carried by using experimental animal model. In general bulk-forming laxatives
which are also called ‘fiber supplements’ are the gentlest on your body and
safe to use long term. Metamual and Citrucel fall into this category (Stacewicz et al., 2001). Stimulant laxatives are the
harshest and shouldn’t be used long-term. Oral laxative may interfere with the
absorption of some medications and food nutrients. Also rectal laxatives do not
have this effect. And both oral and rectal laxatives can lead to an electrolyte
imbalance especially after a prolong use (Lederle et al., 1990).
Antioxidants comprises of a range of substances that play a role in
protecting biological system against the deleterious effect of oxidative
processes on macromolecules such as proteins, lipids and carbohydrate, many of
those substances which contribute to the prevention and treatment of diseases
in which reactive species of oxygen are involved are natural molecules of
plants. Generation of free radicals or reactive oxygen species during
metabolism and other activities beyond the antioxidants capacity of a
biological system gives rise to oxidative stress (Zima et al., 2001). Thus, the antioxidant statue in human reflects the
dynamic balance between the antioxidant defense and pro-oxidant conditions and
this have been suggested as a useful tool in estimating the risk of oxidative
damage (Tiwari, 2004).
1.2
JUSTIFICATION FOR THE STUDY
Antimicrobial diseases still remain
one of the major causes of death in developing countries, where there is
inadequate production of pharmaceutical drugs, the use of plants that are
inexpensive and easily available is Synedrella
nodiflora It has been revealed by many herbal practitioners to cure
constipation, diarrhea, dysentery, infections, pains etc. but there was no
scientific prove. Based on this background, the present study was intended to
obtain information of the antioxidant, laxative and antimicrobial activities of
Synedrella nodiflora leave.
1.3
AIM AND OBJECTIVES
AIM;
Ø To
determine the antioxidant, antimicrobial and laxative activities of ethanol
leave extract of Synedrella nodiflora on
experimental animal model and 6 bacterial strains bacteria.
OBJECTIVES
Ø To investigate the laxative effect of ethanol,
leave extract of synedrella nodiflora
on experimental animal model.
Ø To
determine the antioxidant activity of leave ethanol extract of synedrella nodiflora.
Ø To
determine the antimicrobial activities of ethanol, leave extract of synedrella nodiflora on some pathogens
microbes.
1.4
DEFINITIONS OF TERMS
According to the dictionary of
Biochemistry 2nd edition, Wiley publication New York and the
advanced English dictionary they following are defined.
Antioxidant;
These are substances that inhibits
oxidation reactions promoted by oxygen or peroxides. In order words, they act
or have agents that act against oxidation.
Toxicity;
This is the degree of harmfulness of a substance
for an organism, they capacity of a substance to produce injury.
LD50;
The amount of a substance required to kill
half a given population, thus a measure of toxicity (acute).
Laxatives;
These are foods or drugs that causes or
help the bowel to empty.
M.I.C;
the minimum inhibitory concentration is
defined as the lowest concentration able to inhibit any visible microbes’ growth
on the culture plates.
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