antibacterial activity of malay traditional plants(4 draft jmpr)
DESCRIPTION
Antibacterial Activity of Malay Traditional PlantsTRANSCRIPT
Antibacterial Activity of Selected Edible Medicinal Plants
Mohamad Zainuddin, Mohamad Farhan, Abdul Hamid, Azizah1,2 h* and Khatib Alfi
1 Department of Foood Science, Faculty of Food Science and Technology, Universiti Putra
Malaysia, Serdang, Selangor, Malaysia
2 Agrobiotechnology Institute, Ministry of Science and Technology, Malaysia.
*Corresponding author. Tel:+603-89468374; Fax: +603-89423552
Email address: [email protected]; [email protected]
Abtract
Seventeen edible traditional medicinal plants that are usually being used as salads and
traditional medicine in Malaysia were investigated for their antibacterial properties in vitro.
Antibacterial activity of methanol extracts of the each plants wereas evaluated by using disc
diffusion and agar dilution methods against pathogenic strains of Gram positive (Bacillus
cereus, Listeria monocytogenes and Staphylococcus aureus, Bacillus cereus and Listeria
monocytogenes) and Gram negative bacteria (Escherichia coli and Salmonella typhimuri and
Escherichia coli ). Results from disk diffusion assay showed that nine9 plants had appreciable
antibacterial activity including Andrographis paniculata, Borsenbergia rotunda, Cosmos
caudatus, Curcuma Xanthorhiza, Kaempheria galanga, Lawsonia inermis, Melicipe lunu,
Muraya koenigii and Cosmos caudatus, Piper betle, Melicipe lunu, Lawsonia inermis,
Borsenbergia rotunda, Andrographis paniculata, Muraya koenigii and Kaempheria galanga.
While ten plantsin agar dilution assay showed 10 plants active by including Pipper longum
exhibited antibacterial activity in agar dilution assay. Each of the active plants inhibited at
least one strain bacteria with minimum inhibition concentration (MIC) range between 1-32
1
1
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
mg/mL. Interestinglly, C. xXanthorhiza showed the strongest antibacterial activity followed
by C.caudatus, P.betle, M.lunu, L. inermis, B. rotunda, A. paniculata, M. koenigii, K. galanga
and finally P.longum. While L.inermis and P.betle showed broadest activity against all
bacteria tested. The rest of the plantsamples such as Centella asiatica, Gynura procumben,
Justicia gendarussa, Morinda cintrifolia, Psophocapus tetragonolobus, Sesbania grandifolia
and Talinum triangulare did not revealshow any antibacterial activity. It can be concluded
that a number ofcertain edible traditional medicinal plant extracts possesed antibacterial
activity and theseit may serve as new sources of antibacterial agents.
Keywords: Malaysian edible medicinal plants; Foodborne pathogen; Antibacterial activity;
Disc diffusion method; Agar dilution method
1.Introduction
Food borne diseases areis still a concern for both consumers and food industry despite
the use of various preservation methods. Food safety researchers and regulatory agencies are
continuously concerned with the high and growing number of illness and outbreaks caused by
some pathogenic and spoilage microorganisms in foods (Shan et al.,2007). It ishad been
estimated that 76 million people in United States had suffered from foodborne illness each
year (Mead et al., 1999). It WHO (2007) also reported that 30% of people in industrialized
countries suffer from food borne diseases each year and in 2005 alone, at least 1.8 million
people died form diarrhea related diseases worldwide (WHO, 2007). Many countries losee
2
2
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
billions of dollar due to medical cost, productivity losses and value permature deaths related
to food borne diseases (Snowdon et al., 2002).
Foodborne diseases can be divided into two groups: which are food infection and food
intoxication. Foodborne pathogen is the main role cause of disease which many occur as a
result of cross contamination, improper handling of food and temperature abuse., it happens
via cross contamination, improper handling and temperature abuse. Food intoxication
occursn happened when the patient consume food that contains harzadous toxic chemicals
produced by bacteria such as Salmonella sp. and Compylobacter (Melzer and Shah, 2009).
This can It also can happen even if the toxin producing microorganisms that produced the
toxin is are no longer present or unable to cause infections. On the other hande, food infection
iwas caused by the presencet of infectious pathogens in the consumption foods. Consumption
of contaminated foods will result in multiplication of microorganisms in the intestine, with
release toxins which invade and damage the epithelium cells. consumed that will multiply in
the intestine and release their toxins which invade and damage the epithelium cells.
Salmonella and Compylobacter sp. had been reported to beas one of the common bacteria
involved in food infections. Others While thereported microorgaismsthat implicated less
frequently in food borne infectionus arewere Listeria monocytogenes, Escherichia coli,
Bacillus cereus and Streptococcus sp. (Taylor., 2002).
In an effort toorder to control microbial growth and reduce the incidence of food
poisoning and spoilage, many synthetic antimicrobials have been usedproduced (Shan et al.,
2008). AlEven though synthetic perservatives are effective, there is still concerns regarding
consumers are still worried about their potential toxicity (Tang et al., 2008). They have also It
3
3
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
also had been reported to be assosiated with causes many side effects such as sleepinessy,
halucination, fever, paranoriaid and amenisia (Willey et al., 2008). In addition, synthetic
drugs also haves been implicatedrelated with the evolution of drug resistant microorganism.
With the increased awareness of people about these health risks, the consumption of raw
vegetables and fruits has increased significantly as individuals have become more heatlh-
conscious and aware of the importance of plant-based diets in combating the onset of such
diseases (Shuib et al., 2010).
Due to all these increasing drwabacks, many researchers have attempted to find new and safer
sources of antimicrobialagents from natural sourcesthe problems, mmany researcherstend to
go back to natural product as an alternative way in finding a new source of antimicrobial
agent. Each Pplants possesed their own characteristic of self defence mechanism against
viruses, bacteria, fungals, bugs and herbivor animals. From the characteristic of the selected
plants, the ancient people had tested those plants until they found which plant is safe and
effectively cure specific illnesses. Those knowledges that had been claimed to be safe had
been passed down from generation to generation. Today, . Nnumerous traditional medicinal
plants have been explored for in more detail to find their bioactivity potential. such
Antioxidant activity have been found in Cosmos caudatus, Murraya koenigii (huda et al.,
2009) as antioxidant, anti diabetic activity in Gynura procumbens (zurina et al.,2010),
antimicrobial activity in Morinda elliptica, Borreria latifolia, Sida rhombifolia (ali et al.,
1995), anticancer activity in Jatropha curcas (Ehsan et al., 2011), anti inflammatory activity
in Piper sarmentosum, Psophocarpus tetragonolobus, Sauropus androgynus (Lee et al.,
2011) and othersmany more.
4
4
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
Those plants also were cheap to buy compared to the synthetic drugs and some
consumers self planted the plants which seem easy to grow. This fact had been supported
from Eloff (1998) where in his report, in 1998, about 80% of the people in developing
countries almost exclusively use traditional medicinal plants.
NumerousSome traditional medicinal plants arecan be consumed as ingredient in
cooking or eaten it rawly as salad. There is common belief Most people highly believe that
those plants are safe for consumption as they have been consumed from generation to
generation without any toxicity reportto consume since there is no complain about the
poisoness or toxicity report of the plant from generation to generation. Most of the edible
traditional plants have many medicinal effects in our digestion system such as for treating
stomach diseases, diarrhea, dysentry and diuretic (Sankaranarayanan et al., 2010; Yasni et al.,
1994; Chowdhury et al., 2008; Gowril and Vasantha., 2010). Plants have also been used to
treat outer body conditions It also had been reported have outer body treatment such as skin
diseases, itches, wounds, bruises, irritant, boils and ulcers. (Chaudhary et al., 2010; Shaari et
al., 2006; Ridtitid et al., 2008;Manoj et al., 2004; Perry, 1980). All those therapeutic values
are related towith the presence of phytochemical compounds phytochemical compounds
(flavonoids, isofalavones, lignans, cinnamic acids derivatives, steroids, carotenoids,
terpenoids, etc), vitamin, polysaccharides, proteins and minerals presentcontain in the plants.
These phytochemicals may also have antibacterial activity. Therfore objectives of this study
weare to screen antibacterial activity of seventeen edible medicinal plants by using different
method approaches and evaluate the lowest concentration required for the screen and evaluate
the potential of antibacterial activity against different strains of food borne bacteria. of
selected edible traditional Pplants were seleceted based on their traditional medicinal practice
used against different strains of food borne bacteria.
5
5
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
The sample plants for this research is Andrographis paniculata, Boesenbergia
rotunda, centella asiatica, cosmos caudatus, curcuma xanthorhiza, gynura procumbens,
justicia gendarussa, kaempferia galangal, Lowsonia inermis, Melicipe lunu, Molinda
cintrifolia, Muraya koenigii, Pipper betle, Piper longum, Premna Cordifolia, Psophocapus
tetragonolobus, Sesbania grandifolia, Talinum triangulare and Vitex nogundo. These samples
were selected based on their used as salad, food ingredient and contribution in medicinal
properties. Table 1.1 show the ethnomedicinal uses of selected plants in this study.
Since in 1550 BC, Egyptians had been used spices and herbs such as Cinnamon, cumin and
thyme as a food perservation and mummification (Webb & tanner, 1994 ; Hirasa & takemasa,
1998). Until now numerous studies have been published on the antimicrobial activities of
plants extracts against different types of bacteria (Shan et al.,2007; Cos et al., 2002; Kumar et
la., 2006; Vivek et al., 2008; Buwa and Van., 2006; Agnihotri et al.,2008; Khan and
Omoloso.,2008; Tadhani and Subhash.,2006;Tsai et al.,2008 and Wang et al.,2008). However,
only a few studies focused on the potential of consumption vegetables plants as sources of
antibacterial compounds that could inhibit foodborne microorganisms. The objective of this
study are to determine the antibacterial activity of extracts from “salad” with traditional
medicinal properties against food borne microorgansims
Table 1.1 Ethnomedicine uses of selected consumption plants “salad” in Malay communtiy.
Species name Common name Part tested Ethnomedicinal uses
Pipper betle
Sireh Leaves
6
6
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
Lowsonia
inermis Inai
leaves
Melicipe lunu T.Burung leaves
Borsenbergia
rotunda
T.kunci tuber
curcuma
xanthorhiza T.lawak
tuber
Cosmos
caudatus
U.raja
leaves
Andrographis
paniculata
H.bumi leaves
Muraya
koenigii Kari
leaves
kaempferia
galangal Cekur
leaves
Piper longum Kaduk leaves
Talinum
triangulare
K.Belanda leaves
7
7
Sesbania
grandifolia Turi
leaves (.,
Psophocapus
tetragonolobu
s
K.botol
pods
Molinda
cintrifolia Mengkudu
leaves
gynura
procumben S.nyawa
leaves
justicia
gendarussa G.rusa
leaves
Centella
asiatica Pegaga
leaves
2. Materials and methods
2.0 materials and chemicals/reagents
8
8
146
147
148
149
150
Traditional Malaysian medicinal plantsplants like Centella asiatica, Cosmos caudatus,
Curcuma xanthorhiza, Justica gendarussa, Lawsonia inermis, Kaemperia galangal, Melicipe
lunu, Morinda cintrifolia, Muraya koenigii, Piper betle, Pipper longum, Psophocapus
tetragonolobus, Sesbania grandifolia, Talinum triangulare, Gynura procumbens,
Borsenbergia rotunda and Andrographis paniculata were selected based on their traditional
medicinal uses asare listed in Table 1. The plant parts were collected from the Traditional
Medicine Plant Plot, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Dimethyl
sulfoxide from Fisher Scientific (Leicestershire, UK),. Chloramphenicol (30ug) and
Tetracycline (10ug), Mueller Hilton Agar (MHA) medium from Oxoid ( (Hampshire,
England) and NuncoTM Surface (Rosklide, Denmark)., respectively.
2.3 Microbial strains and culture
Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 14579, Listeria
monocytogenes ATCC 19115, Campylobacter jejuni ATCC 29428, Salmonella typhimurium
ATCC 13311 and Escherichia coli ATCC 25922 were purchased from ATCC (American
Type and Collection Centre). Each bacterial strain was cultivated in Nutrient Agar and
incubated for 18-20 hours at 37◦C in an incubator (Memmert, Germany). Then three or five
colonies bateria of the same morphological type form on agar plate were then taken up and
suspended in 10mLl sterile saline followed by then vortexingxed thoroughly. The bacterial
suspension was prepared according to was followed 0.5 Mcfarland standards. The inocoloums
were used within less than 15 minutes ofafter preparation.
2.2 Preparation of samples
plant partscollected from the Traditional Medicine Plant Plot, Universiti Putra
Malaysia, Serdang, Selangor, Malaysia. Plant materials obtained were washed underwith
9
9
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
running tap water, before being chopped into pieces, followed by drying then dried in
conventional oven (Memmert, Germany) at 45oC for two days. Dried plants were then and
then grounded into powder form and kept at. The powders were then kept at -20oC until
further analysis..
2.4 Preparation of crude extract
100g of sample The sample was mixed with absolute analytical methanol (Merck,
Germany) at 1:10 ratio and leftaved in shaking water bath for 24 hours at 45◦C. Samples were
then filtered using whatman filter paper No.1 and the solvent was removed by using Eyela
rotary evaporator (Tokyo Rikikai Co. Ltd, Japan) set at 40◦C.
2.5 Antibacterial screening assay
2.5.1 Disk diffusion method
The cultures were prepared according to Institute Cclinical Laboratory Standard
(2001) standardization and the antibacterial assay was prepared according to Lee et al., (2007)
with some modifications.
Sterile cotton was dipped into the inoculum and then rotated firmly to remove the excess
liquid, followed by streaking into thethen streaked into entire surface of Muller Hilton Agar .
This was repeated 3 times at 60 degree rotation in ensuring for three times. Each swab was
applied in 60 degree rotation to make sure all inoculums were well distributed and finally
swab all around the edges of the agar surface. Twenty microlitter of the extracts
(40mg/mL)was impregnated into 6mm diameter paper disc.The extracts were diluted in
Dimethyl sulfoxide (DMSO) and adjusted to 40mg/mL concentration. Twenty microlitter of
the solution was dispensed into 6mm diameter paper disc. The paper discs were left 30
minutes to dry, then placed the disc on the surface of Muller Hilton Agar with inoculum on it.
10
10
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
The plates were then incubated in an inverted position at 37 ◦C in an incubator for 18-20
hours. Tetracycline was used as a positive control, and the . While for negative controlblank
consisted of was just only Nutrient agar. The antimicrobial activity was assessed based on the
measurement of the diameter of the clear zone around the paper discs. In this study, tThree
replicates carried outwere prepared for each type of bacteria tested.in this study.
2.5.2 Agar Dilution method.
The AAgar Dilution method was conducted according to the method proposed by
Jennifer, (2001) with some modifications. Crude extract (64mg)Sixteen four mg crude extract
was diluted with 10% DMSO and used as stock solutiiton for further experiment. Form Tthe
stock solution, (6.25mL) was taken out and mixed with 3.75mL sterile Muller Hilton agar
(MHA) in sterile round plate (90 x 15mm) which given a final concentration of 40mg/mL.
The temperature of MHA was then brought to 50 °C before pouringed to the plate. Then
swirled until thoroughly mixed. The plates were then left for 10 minutes and allowed to
solidify. Then 1 uL of suspensions from the prepared inoculums were placed on the agar
surface. Chloramphenicol was used as a positive control. While for Tthe blank consisted of
was MHA only only. The plates were then incubated in an inverted position at 37◦C in an
incubator (Memmert, Germany) for 18-20 hours. The activity was then documentedwill be
noticed based on visualization, whether either the bacteria is ablecapable to growt h or not.
Three replicates were carried atprepared for each type of bacteria tested in theis study.
2.6 Determination of Minimum Inhibition Concentration by using Agar Dilution
method.
The method was theere same aswith the previous agar dilution method with the
exception of. Only the concentaration of the sample usedadjusted. The suitable amount of
11
11
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
extract was added on to the plate (90 x 15mm) followed by then mixinged with a specific
setence volume of sterile Muller Hilton Agar (MHA) to produce final concentration of 0.5,
1.0, 2.0, 4.0, 8.0, 16.0 and 32.0 mg/mL. Chlorampehenicol (30ug) was used as a positive
control. Two replicates were prepared for each type of bacteria in this study.
3. Result
In the present study, 17 methanol extracts of selected tropical plant collected from
Taman Pertanian Putra (TPU), Universiti Putra Malaysia (UPM), Serdang, Selangor,
Malaysia were tested against 3 gGram-negative (Bacillus cereus, Listeria monocytogenes,
Staphylococcus aureus) and 2 gram-positive (Salmonella thyphii, Escherichia coli)
foodborne bacteria by using on the basis of disc-diffusion and agar dilution assay. Table 3.1
showed the screening results of antibacterial activity of plants extracts by using Disc-diffusion
method and Agar dilution methods. While mMinimium inhibition concentration (MIC)
obtained from were examined by using Agar Dilution assay isas showed in table 3.2. The
Rresults revealedshow that only 10 out of 17 plant extracts exhibitedshowed potent
antibacterial activity which inhibited at least one selected bacteria strain.
quantitaively assesed by the presence or absence of inhibition zones and zone diameters
(Table 1), MIC values (Table 2).
.
Results screening and mic
The Sscreening results from disk diffusion method showed that 9 out of 17
plants posessed potent antibacterial activity where atleast inhibiting one bacterium tested in
this study. Pipper betle and lawsonia inermis extract showed broadest anti-bacterial activity
against all tested bacteria as compared to the other plant extracts. where They also showed the
highest antibacterial activity in this screening result where the extracts wereit is capable to
12
12
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
inhibiting more than 10mm diameter inhibition zone atleast for two selected bacteria. For
Melicope lunu, Curcuma xanthorhiza, Cosmos caudatus and Andrographis paniculata assed
in the present study showed moderate antibacterial activity where they it inhibiteds all the
gram positive bacteria strain (S. aureus, B. cereus and L.monocytogenes) withhich diameter
inhibition zone in the range from 8.0 to 10.0 mm. Low antibacterial activity was possesed by
Kaempferia galanga, Boesenbergia rotunda and muraya koenigii showed low antibacterial
activity with inhibition of atleast one bacteria and diameter inhibition zone of 6.0 – 8.0
mmwhich capable to inhibit atleast one bacterium strain where diameter inhibition zone show
in the range of 6.0 to 8.0 mm. The Oother plant extracts such as Centella asiatica, Justica
gendarussa, Morinda cintrifolia, Pipper longum, Psophocapus tetragonolobus, Sesbania
grandifolia, Talinum triangulare and Gynura procumbens did not show any antibacterial
activity in the present study.
Mean while screening byResults obtained from the Agar Agar Dilution method
revealed that 10 out of 17 plant extracts (40mg/mL) possesed antibacterial activity against at
least 1 bacterial strain at the same concentrations as in disk diffusion assay (40mg/mL). As
expected, Piper betle, Lawsonia inermis and Melicope lunu showed the broadest spectrum of
action against bacteria, inhibiting all the strains tested. Borsenbergia rotunda also showed
quite broad activity where it could inhibit 4 out of 5 strains tested. Curcuma xanthorhiza,
Cosmos caudatus, Androphanis paniculata, Kaempferia galangal and Muraya koenigii also
possesed antibacterial activity against all gram positive strains. The lowest activity was shown
by Piper longum where it only inhibited Listeria monocytogenes. While the other plant
extracts such as Centella asiatica, Justica gendarussa, Morinda cintrifolia, Psophocapus
tetragonolobus, Sesbania grandifolia, Talinum triangulare and Gynura procumbens samples
did not show any antibacterial activity.
13
13
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
Plant extracts showing potentialBased on Screening activity, only those who have the
positive antibacterial activity were then further tested is using the have been proceeded to
further study which is evaluating the Minimum Inhibition Concentration (MIC), assay which
assesses the of selected sampes. MIC is a minimum concentration requiredneeded to inhibit
bacterial growth. The results showed that Piper betle, Cosmos caudatus, Melicipe lunu and
Curcuma xanthorhiza showed high antibacterial activity against gram positive bacteria, where
it inhibited the bacteria at low concentration (1.0 – 2.0 mg/mL). Melicipe lunu, Borsenbergia
rotunda and Andrographis paniculata showed modest activity where inhibition occurred at a
concentration of 4.0 to 8.0 mg/mL respectively. Muraya koenigii, Kaempferia galanga and
Piper longum showed low activity in this study where it required high concentrations of
extract, in the range of 16.0 to 32.0 mg/mL. The highest activity was shown by Curcuma
xantthorhiza where it inhibited all the bacteria tested at a low concnetration of 1 mg/mL. On
the other hand, gram-negative bacteria were only affected by Pipper betle, Lawsonia inermis,
Melicipe lunu and Borsenbergia rotunda. Piper betle exhibited high antibacterial activity
where it inhibited the bacteria tested at 2mg/mL. Followed by Lawsonia inermis which
exhibited modest activity ( 4.0 – 8.0 mg/mL). Lastly, Melicipe lunu and Borsenbergia
rotunda showed low antibacteria activity where the MIC value for the bacteria tested were the
range of 16 to 32 mg/mL. Piper betle, Lawsonia inermis, Melicope lunu and Borsenbergia
rotunda had the broad spectrum of activity where it could inhibit almost all the bacteria tested
which MIC value in the range of 32mg/mL to 1mg/mL. While, Curcuma xanthorhiza,
Cosmos caudatus, Androphanis paniculata and Kaempferia galangal showed modest range
which it only inhibited 3 out of 5 bacteria tested. The lowest antibacterial effect was Piper
longum, it only inhibited 1 bacterium which is Listeria monocytogenes .
14
14
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
From the bacteria testedThe study revealed that, Esherichia coli showed thewere most
resistant and exhibit only slight susceptibility to Piper betle, Lawsonia inermis and Melicope
lunu with MIC value ranginge from 2 to 32 mg/mL. The most susceptible bacteria was
Listeria monocytogenes, which was inhibited by 10 plant extracts with MIC value range from
1 to 32 mg/mL. Negative control was observed that no inhibition of the strain to growth. It
can be said that the studied plant extracts were less susceptible to the gram negative bacteria
than gram positive bacteria.
4. Discussion
Ten out of 17 plants used From 17 plants that been used as salad and herbs in
traditional medicine in Malaysia in the present study were found to , ten of them exhibited
potent antibacterial activities against six foodborne bacteria. This study also revealed the
antibacterial activity were to same extent assay dependent. Different observation were recrded
for disk diffusion and agar dilution assay. This is supported by previous literature (rios et al.,
1980), who the different screening results between disk diffusion and agar dilution assay. The
different had been expected as descibed by Rios et al., (1988), where the author reportedstated
that agar diffusion method was only suitable for testingstudying polar compounds since the
polar compounds are capable to disolve and diffuse into the agar. On the other hand While
non-polar compounds were unable to diffuse into the agar and might end up with false result.
However, thisthis method is widelyhad been well used by numerous reseachers as a
preliminary study since it iwas affordablecheap, easy and fastless time consuming. On the
other hand, agar dilution method is applicable to both polar and non-polar compounds since
the microorganisms are can directly in contact with the compound that had been mixed with
the agar. However, the Even do this method iswas not very populor due to its being time
consuming and utilizing among researchers since it high amount of quite difficult and need a
15
15
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
lots of crude extracts. Agar dilution method has beenwas claimed to be a suitable method in
evaluating antimicrobial potential of for complex samplesextract such as plant extracts (Rios
et al., 1988). This could explain the different results on antibacterial activity screening of
M.lunu, B.rotunda, K.galanga and P.longum extract in this study.. Therefore from the
screening results, the samples that have shown antibacterial activity in screening had been
carried out to determine their MIC by using agar dilution method. Rios et al., (1988) also
reported in his review that agar dilution results were relevant for MIC value.
Antibacterial activity demostrated by plant extracts is usually attributed to an array of
phytochemicals present in the Based on the literature review, bioactive compounds of
Melicope .lunu, Kaempheria .galanga and P.iper longum had been reported to contain
nemurous of bioactivenon-polar compounds including. For instance, M.lunu possesed
limonene and, terpene and α-ocimene (Goh et al., 1995), While K.galanga contained
consisted ofcavene, cineol, ethyl cinnamate and coumarin (Othman et al., 2006; Ismail 2000).
P.longum is richcontained peperine, assarinin, piperidine and sarmintine (Majumdar et al.,
2002; Sawangjaroen., 2005; Selvendiran et al., 2005; Nalin and Rahim., 2007). All these
bioactive compounds have been reported to exhibit potent antibacterial activity (Cowan.,
1999; Sibel.,2003; Iqbal et al.,2006) In addition,While B.rotunda hasve been reported to
contain cChalcones a non which are non polar flavanoid derivatives, as a one of it major
compound (Nor AY et al., 2010). P.betle and L.inermis may have high amount of polar and
non-polar compounds, which could be the reason that explained the most active antibacterial
results in disk diffusion assay. P.betle had been reported to havesimilarily consisted of high
phenolic compounds such as terpene , pyrocatechin, cavicol, cavibetol, carvacrol, eugenol and
allilpyrocatechol (Farnsworth and Bunyapraphatsara, 1992), alkaloids, saponins and tannins
(Anonymous., 1992) thatwhich are known to have very potent antibacterial agent. Similar
16
16
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
bioactive compounds werealso had been found in L. inermis in addition towith some other
compounds such as flavanoids, terpenoids, quinones, coumarins, xanthones and lawsone
(Mikhael et al., 2004; edrini et al., 2002; Chaudary et al., 2010). In the present study,
C.xanthorhiza and C.cosdautus were expected possessed a lot of essential oil compound
where Vimala et al. (2003) reportedfound that C.caudatus containing stigmasterol,
ssquiterpene and hydroxy eugenols and. Similarly, C.Xanthorhiza has been reported to
contain eugenol (Ruslay et al., 2007; Noraida 2005), terpinene and xanthorhizol (Chatterjee et
al., 1999) where theose essential oils have been reported to exhibitcointain potent antibacterial
activity. Andrographis .paniculata and M.koenigii showed only moderate antibacterial
activity, and is probaly due to the lower amount of maybe it is because these plants have less
amount of antibacterial bioactive compounds with antibacterial activity. Several studies had
found that A.paniculata contained compound such as diterpenoids, eugenol, caffeic acid,
tritriacetone and flavonoids (Joganath et al., 2000; Sukrasno et al., 2011).. While, M.koenigii
had been found to contain caumarins, terpenoids, murrayazoline and sesquiterpenes (Noraida
2005; Yadaz et al., 2002 and Tachibana et al., 2001).
Discn mode of action
The antibacterialmicrobial activities of phytochemicals may involve multiple modes of
action as been described by Cowan (1999). For example, Pphenolics are a broad class of
compounds that have a variety of antibacterial mechanisms. Each of the subclasses have their
own specific mehanisms of action. Simple phenols such as catechol and epicatechin impart
their antibacterial activitywork by substrate deprivation and membrane disruption of
bacteria`s cell wall. Phenolic acids and quinones act by binding to and form adhesins
complexing with cell wall and deactivated the enzymes. Similarly, flavonoids binds to
adhesins and form a complex with cell wall. Tannins react by binding to protein and adhesin,
17
17
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
distrupting membranes, forming complex with the cell wall, inhibiting enzymes and caused
subtrate deprivation. Other classes of phytochemical compounds such as terpenoids
(capsaicin) and essential oils act by membrane distruption and alkaloid act by intercalating
into the cell wall or DNA. However Shan et al., (2007) reported that there are many other
compounds and reactions that may had contributed before the final mechanisms of
antibacterial activity take it place.
Since there are many possibilities of antibacterial mechanisms, all of these mechanism are not
separate targets. Some of the mechanism of inhibition are effected as a consequense of
another mechanism being targeted (Shan et al., 2007). However the mechanism of
antimicrobial agents are also depend on the type of microorganism and mainly related to their
cell wall structure and outer membrane arrangement
Among the bacteria tested, Escherichia coli and Salmonella thyphii werewas found to
be the most resistant strain, that was only inhibited to some extent by . Only P.betle, L.inermis
and M.lunu extracts were found able to inhibit this bacterium. This observation was agreed
with that of previous studies that gGram-negative bacterial generally were more resistant to
traditional herbs extract as compared to that gram positive bacteria (Shan et al., 2007).
Numerous In fact, numerous studies have alsobeen reported that gGram-negative bacteria
were becoming more resistant towards many available antibiotics current available in the
market (Alonso et al.,2000; Sader et al., 2002). This is might be explained due to the
differences in morphological constitutions between the microorganisms. Gram-negative
bacteria possess an outer membrane and unique periplasmic space which is lacking inere
gram–positive bacteria was lacking does not ( Nikaido, 1996; Duffy and Power, 2001). The
outer membrane of gram-negative bacteria is covered by hydrophilic surface and rich in
lipopolysacride molecules that function as a barrier to the penetration of any harmful
subtances while the periplasmic space contains several degradative enzymes which are
18
18
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
capable ofin breaking down the introduced molecules from outside (Russell, 1991; Nikaido,
1994;Gao et al., 1999). GThe Gram- positive bacteria on the other hand, does not has an outer
membrane, but onlyand the cell wall structure consisting of only having peptidoglycan layer
thatwhich is not an effective permeability layer barrier. Thus, antibacterial chemical subtances
can easily penetrate into the bacteria cell wall and cyctoplasmic membrane, resultinged in a
leakage of the cyctoplasm and it coagulatione (Kalemba and Kunicka, 2003). AlEventhough,
the cell walls of gram- negatives are more complex than gram-positive bacteria (Nostro et
la.,2000; Hodges, 2002), results from but in the present study, revealed that some of the
extracts can have still exert ed some degree of inhibition against selected the gram negative
bacteria .
It can be said that the studied plant extracts were less susceptible to the gram negative
bacteria than gram positive bacteria.
5. Conclusion
In Extracts of Piper betle, Lawsonia Inermis, Melicope lunu, Borsenbergia rotunda,
Curcuma xanthorhiza, Cosmos caudatus, Andrographis paniculata, Muraya koenigii ,
kaempferia galangal and Pipper longum demostrated activity against at least one strain of
bacteria was inhibited. The nemurous and different phytochemical compounds in these plants
may be responsible for their varying nemurous applicable in due to antibacterial activities.
This study has provided on sight on the potential use of certain Malaysian medicinal plants as
antibacterial agents. However further studies are required to identify bioactive compound of
interest.
nalternative and in growth
19
19
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
20
20
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
4. Discussion.
5.Conclusion
Table
Table 1: Ethnomedicine uses of selected edible traditional medicinal plants in Malaysia
communtiy.
Species name Common name Part tested Ethnomedicinal uses
Pipper betle Sireh Leaves Digestive, stimulative, carminative andaphrodisiac (Sankaranarayanan et al., 2010).
Lowsonia inermis Inai Leaves
Alleviating jaundice, skin diseases, venereal diseases, smallpox andspermatorrhoea (Chaudhary et al., 2010)
Melicipe lunu Tenggek Burung Leaves Treatment of itches and wounds (Shaari et al., 2006),
Borsenbergia rotunda Temu Kunci Rhizom
Ailment, illness and confinement. Rhizomes are also taken as carminatives for relieving flatulence.(Chan et al., 2008 )
Curcuma xanthorhiza Temu Lawak Rhizom
Treatment of stomach diseases, liver disorders, constipation, bloody diarrhea, dysentery, fever in children, hemorrhoids, and skin eruptions (Yasni et al. 1994)
Cosmos caudatus Ulam Raja Leaves
Blood cleansing, induction of uterine contractions and prevention or cure of ailments such as diabetes, high bloodpressure, cardiovascular disease, arthritis, fever and coughs (Abas et al., 2006)
Andrographis paniculata Hempedu Bumi Leaves
Treatment of upper GI tract and upperrespiratory infections, fever, herpes and other chronic diseases. (Roy et al., 2010)
Muraya koenigii Kari Leaves
Relieve nausea, indigestion, vomiting; treatment of diarrhea and dysentery (Chowdhury et al., 2008).
Kaempferia galangal Cekur Leaves
Treatment of Tinea versicolor, and eye diseases and seizures, rheumatism, asthma, headaches, cough, toothaches, bruises and wounds (Ridtitid et al., 2008)
Piper longum Kaduk LeavesTreatment of respirotory tract, cough, bronchitis, irritant, inflammation. (Manoj et al., 2004)
Talinum triangulare Kerekot Belanda Leaves Treatment of diuretic, gastro-intestinal
disorder.(Mensah Et al., 2008)
21
21
451
452
453
454
455
456
457
458
459
Sesbania grandifolia Turi Leaves
Aperient, diuretic, and tonic anddisinfect the mouth and throat (gowri1 and Vasantha., 2010)
Psophocapus tetragonolobus Kacang Botol Pods Treatment of skin sores such as boils and ulcers
(Perry, 1980). Molinda citrifolia Mengkudu Leaves Relief joint pain (Rout et al., 2009)
Gynura procumben Sambung Nyawa Leaves Treatment of malaria, general febrifuge, and
analgesic (Scott., 2006)
Justicia gendarussa Ganda Rusa Leaves
Treatment of fever, hemiplegia, rheumatism, arthritis, muscle pain, lumbago, headache and earache (Ahmad and Holdworth 2003; Anonymous 1959)
Centella asiatica Pegaga Leaves
Against conjunctivitis and other eye injury, wound healing but especially for the treatment of skin diseases such as eczema, leprosy and psoriasis. Treatment of burns, itching and insect bites (Gupta et al., 1999; Zainol et al., 2008)
22
22
460
461
462
463
464
465
466
467
468
469
470
471
472
473
474
475
476
3.1 Disk diffusion method
Table 3.1: Antibacterial activity of plant extracts screening of 40mg/mL concetration by using
disk diffusion and agar dilution method.
SamplesPlant extract
Disk diffusion method Agar dilution methodDiameter of zone inhibition(mm) Inhibition of bacteria growth
Gram-positve bacteria Gram-negative bacteria
Gram-positive bacteria
Gram-negative bacteria
BC SA LM EC ST BC SA LM EC ST
P. betle 7.7 ± 0.3
11.8 ± 0.6
7.3 ± 0.3
15.4± 1.7
10.4 ± 0.1 + + + + +
L. inermis 14.7 ± 0.3
8.4 ± 0.5
14.8 ± 0.5
7.2 ± 0.1
7.0 ± 0.1 + + + + +
M. lunu 9 .2 ± 0.3
9.0 ± 0.5
9.5± 0.4 - - + + + + +
B. rotunda 7. 2 ± 0.2 - 7.1±
0.1 - - + + + - +
C. xanthorhiza
8.0 ± 0.2
7.8 ± 0.3
7.8± 0.3 - - + + + - -
C. caudatus 7.9 ± 0.2
8.9 ± 0.4
8.6 ± 0.5 - - + + + - -
A. paniculata 8.0 ± 0.1
8.1± 0.2
7.7 ±0.3 - - + + + - -
M. koenigii 7.1±0.1
7.1±0.2
7.6 ±0.4 - - + + + - -
K. galangal - - 7.2 ±0.3 - - + + + - -
P. longum - - - - - - - + - -T. triangulare - - - - - - - - - -
S. grandifolia - - - - - - - - - -P. tetragonolobus
- - - - - - - - - -
M. cintrifolia - - - - - - - - - -G. procumben - - - - - - - - - -
J. gendarussa - - - - - - - - - -
23
23
477
478
479
480
481
482
483
484
C. asiatica - - - - - - - - - -Tetracyline (30ug)
17.4 ± 1.4
21.3 ± 3.6
19.9 ± 0.7
19.3 ± 1.6
22.4 ±2.3 - - - - -
Chloramphenicol (30ug) - - - - - 4.0
ppm8.0
ppm2.0
ppm8.0
ppm8.0
ppmConcentration used 40mg/mL .(+) positive antibacterial activity (-) no activity; BC= Bacillus
cereus ATCC 14579; SA= Staphylococcus aureus ATCC 25923; LM= Listeria
monocytogenes ATCC 19115; CJ= Campylobacter jejuni ATCC 29428; EC = Escherichia
coli ATCC 25922; ST= Salmonella thyphii ATCC 13311
3.2 Agar dilution method
Table 3.2: Minimum inhibition concentration (MIC) of methanol extract of 17
medicinalsalads plants.
Methanol extract
Minimum inhibition concentration( mg/mL)
Gram-positive bacteria
Gram-negative
bacteria
No. BC SA LM EC ST
1 Pipper betle 1.0 2.0 2.0 2.0 2.0
2 Lowsonia inermis 4.0 4.0 4.0 8.0 4.0
3 Melicipe lunu 2.0 2.0 1.0 32.0 16.0
4 Borsenbergia rotunda 8.0 16.0 8.0 - 32.0
5 Curcuma xanthorhiza 1.0 1.0 1.0 - -
6 Cosmos caudatus 2.0 2.0 1.0 - -
7 Andrographis paniculata 8.0 8.0 8.0 - -
8 Muraya koenigii 16.0 16.0 16.0 - -
9 kaempferia galangal 32.0 32.0 32.0 - -
10 Piper longum - - 32.0 - -
11 Chloramphenicol (ppm) 4.0 x 10- 8.0 x 10-3 2.0 x 10-3 8.0 x 10-3 8.0 x 10-3
24
24
485
486
487
488
489
490
491
492
3 .0
(-) no activity; BC= Bacillus cereus ATCC 14579; SA= Staphylococcus aureus ATCC 25923;
LM= Listeria monocytogenes ATCC 19115; CJ= Campylobacter jejuni ATCC 29428; EC =
Escherichia coli ATCC 25922; ST= Salmonella thyphii ATCC 13311
References
Abdul Manaf A, Saleh HS, Junainah AH,Nor Hadiani I, Nordin JL (1995). antimicrobial
activity of selected malaysian plants. l)ertanikaj. trop. agric. sci. 18(1): 57-61
Ahmad FB, Holdworth DK (2003). Medicinal plants of Sabah, East Malaysia-part 1. Pharm
Biol. 41:340–346.
Ahmad R, Ata A, Islam B, Ashfaq M (1990). Studies on saponins from some indigenous
plants. Pak. Vet. J. 10:146-148.
Alonso R, Fernandez-Aranguiz A, colom K, Cisterna R (2000). Profile of bacterial isolates
and antimicrobial susceptibility: multicenter study using a one-day cut0off. Revista Espanola
de Quimioterapia. 13:384-393.
Anonymous (1959). The Wealth of India. CSIR Publications, New Delhi, pp 312.
25
25
493
494
495
496
497
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
Anonymous (1992). The Wealth of India. The Dictionary of Indian Raw Materials and
Industrial Products. Raw Material, revised (eds). Publication and Information directorate,
CSIR, New Delhi, pp 84-94.
Baily E (1945). Treatment of leprosy, 155: 601.
Beuchat LR, Golden DA (1989). Antimicrobials occurring naturally in foods. Food
Technology, 43:134–142.
Chan EWC, Lim YY, Wong LF, Lianto FS, Wong SK, Lim KK, Joe CE, Lim TY (2008).
Antioxidant and tyrosinase inhibition properties of leaves and rhizomes of ginger species.
Food Chemistry, 109:477–483.
Chao W, Lin B (2010). Isolation and identification of bioactive compounds in Andrographis
paniculata (Chuanxinlian). Chinese Medicine, 5:17.
Chaudhary G, Goyal S, Poonia P (2010). Lawsonia inermis Linnaeus: A
Phytopharmacological Review International Journal of Pharmaceutical Sciences and Drug
Research, 2:91-98.
Chatterjee S, Desai SRP, Thomas P (1999). Effect of γ-irradiation on the antioxidant activity
of turmeric (Curcuma longa L.) extracts. Food Research International, 32:487-490.
26
26
518
519
520
521
522
523
524
525
526
527
528
529
530
531
532
533
534
535
536
537
538
539
540
541
Chowdhury JU, Bhuiyan MNB, Yusuf M (2008). Chemical composition of the leaf essential
oils of Murraya koenigii (L.) Spreng and Murraya paniculata (L.) Jack. Bangladesh J
Pharmacol, 3:59-63.
Cowan MM (1999). Plant products as antimicrobial agents. Clin. Microbiol. Rev, 12:564-582.
Cushnie TPT, Lamb JL (2005). Review: Antimicrobial activity of flavonoids. International
Journal of Antimicrobial Agents, 26:343-356.
Dineshkumar B, Mitra A, Mahadevappa M (2010). Antidiabetic and hypolipidemic effects
of mahanimbine (carbazole alkaloid) from Murraya koenigii (rutaceae) leaves
International Journal of Phytomedicine, 2:22-30.
Duffy CF, Power RF (2001). Antioxidant and antimicrobial properties of some Chinese plant
extracts. International Journal of Antimicrobial Agents, 17:527–529.
Ehsan O, Norhani A, Wan Zuhainis S, Abdul Rahman O, Syahida A, Wen BK, Nor Azlina Z,
Rudi H, Yin WH (2011). Antioxidant, anti-inflammatory and anticancer activities
of methanolic extracts from Jatropha curcas Linn. Journal of Medicinal Plants Research Vol.
5: 49-57.
Eloff JN (1998). A sensitive and quick microplate methods to determine minimum inhibitory
concentration of plant extracts for bacteria. Planta Medica, 64: 711-714.
27
27
542
543
544
545
546
547
548
549
550
551
552
553
554
555
556
557
558
559
560
561
562
563
564
565
Faridah AA, Nordin HLA, Israf DA, Khozirah SA, Umi KY (2006). Antioxidant and nitric
oxide inhibition activities of selected Malay traditional vegetables. Food Chemistry, 95:566–
573.
Farnsworth NR, Bunyapraphatsara N (1992). Thai Medicinal Plants. Medicinal Plant:
Medicinal Plant Information Center, Faculty of Pharmacy, Mahidol University, Thailand. pp 402.
Gao Y, van Belkum MJ, Stiles ME (1999). The outer membrane of Gramnegative bacteria
inhibits antibacterial activity of brochocin-C. Applied and Environmental Microbiology,
65:4329–4333.
Gowri SS, Vasantha K (2010). Antioxidant activity of Sesbania grandiflora (pink variety) L.
pers. International Journal of Engineering Science and Technology, 2:4350-4356.
Gupta AP, Gupta MM, Kumar S (1999). High performance thin layer chromatography of
asiaticoside in Centella asiatica. J. Indian Chem. Soc., 76: 321-322.
Hashim M, Najihah (2005) Chemical Constituents and Biological Activity of Four Melicope
Species (Rutaceace). Masters thesis, Universiti Putra Malaysia.
Hodges N (2002). Pharmaceutical applications of microbiological techniques, In Aulton,
M.E , Pharmaceutics: The science of Dosage Form Design, 2nd (Eds). Harcourt publichers
Limited, London, pp 606.
Iqbal A., Farrukh A., Mohammad O.,(2006). Modern Phytomedicine, Turning medicinal
plants to drugs. WILEY-VCH Verlag GmbH & Co. KgaA. pp 261.
28
28
566
567
568
569
570
571
572
573
574
575
576
577
578
579
580
581
582
583
584
585
586
587
588
589
590
591
Jaganath I.B., Ng LT., Muthuvelu C.,Razali A.R., (2000). Herb, Green Pharmacy of
Malaysia. Vinpress Sdn. Bhd. Malaysian Agriculture Research and Development Institute
(MARDI). pp: 126.
Jennifer MA (2001). Determination of minimum inhibition concentration. Journal of
antimicrobial of chemotherapy, 48:5-16.
Kalemba D, Kunicka A (2003). Antibacterial and antifungal properties of essential oils.
Current Medicinal Chemistry, 10:813–829.
Karapinar M, Aktug SE (1987). Inhibition of foodborne pathogens by thymol, eugenol,
menthol and anethole. International Journal of Food Microbiology, 4:161–166.
Kim YS, Hwang CS, Shin DH (2005). Volatile constituents from the leaves of Polygonum
cuspidatum S. et Z. and their anti-bacterial activities. Food Microbiology 22:139–144.
King SR, Tempesta MS (1994). From shaman to human clinical trials: the role of industry in
ethnobotany, conservation and community reciprocity. Ciba Found. Symp, 185:197–206.
Lee KH, Padzil A, Syahida A, Abdullah N, Zuhainis SW, Maziah M, Sulaiman MR, Israf
DA, Shaari K, Lajis NH (2011). Evaluation of anti-inflammatory, antioxidant and
antinociceptive activities of six Malaysian medicinal plants.Journal of Medicinal Plants
Research Vol. 5(23): 5555-5563
29
29
592
593
594
595
596
597
598
599
600
601
602
603
604
605
606
607
608
609
610
611
612
613
614
615
616
617
Lee S, Chang K, Su M, Huang Y, Jang H (2007). Effects of some Chinese medicinal plant
extracts on five different fungi. Food Control, 18:1547-1554.
Lopez CM, Nitisinpraset S, Wanchaitanawong P, Poovarodom N (2003). Antimicrobial
Activity of Medicinal Plant Extracts against Foodborne Spoilage and Pathogenic
Microorganisms. Kasetsart Journal: Natural Science, 37:460-467.
Machado TD, Leal ICR, Amaral ACF, Dos Santos KRN., Da Silva MG, Kuster RM (2002).
Antimicrobial ellagitannin of Punica granatum fruits.Journal of the Brazilian Chemical
Society, 13:606–610.
Manoj P, Soniya EV, Banerjee NS, Ravichandran (2004). Recent studies on well known
spice, piper longum Linn. Natural product radiance, 3:222-227.
Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV
(1999). Food-Related Illness and Death in the United States. Center for Disease Control and
Prevention, Atlanta, Georgia, USA.
Melzer M., Shah,N (2009). Suspected food poisoning. InnovAiT, 2:80-85.
Mensah JK, Okoli RI, Ohaju-Obodo JO, Eifediyi K (2008). Phytochemical, nutritional and
medical properties of some leafy vegetables consumed by Edo people of Nigeria. African
Journal of Biotechnology, 7:2304-2309.
30
30
618
619
620
621
622
623
624
625
626
627
628
629
630
631
632
633
634
635
636
637
638
639
640
641
Mikhaeil BR, Badria FA., Maatoo GT, Amer MMA (2004). Antioxidant and
immunomudulatory constituents of henna leaves. Zeitschrift fur Naturforschung, 59:468-476.
Nikaido H (1994). Prevention of drug access to bacterial targets-permeability barriers and
active efflux. Science, 264:382–388.
Nikaido H (1996). Outer membrane. In: Neidhardt, F.C. (Eds) Escherichia coli and
Salmonella typhimurium: Cellular and Molecular Biology. American Society for
Microbiology Press, Washington, D.C., pp. 29–47.
Nor Azma Yusuf, Suffian Annuar MM, Norzulaani Khalid, (2011). Rapid micropropagation
of Boesenbergia rotunda (L.) Mansf. Kulturpfl. (a valuable medicinal plant) from shoot bud
explants. African Journal of Biotechnology 10:1194-1199,
Noraida A (2005). Penyembuhan semulajadi dengan herba. Malaysia. PTS Millennia sdn.
Bhd
Nostro A, Germano M P, D`Angelo V, Marino A, Cannatelli MA (2000). Extraction methods
and bioautography for evaluation of medicinal plant antimicrobial activity. Lett. Appl.
Microbiol, 30(5):379-84.
Nychas GJE (1995). Natural antimicrobials from plants. In: Gould, G.W. (Eds.) New Methods
of Food Preservation. Blackie Academic, London, pp. 58–89.
31
31
642
643
644
645
646
647
648
649
650
651
652
653
654
655
656
657
658
659
660
661
662
663
664
665
Perry LM (1980). Medicinal Plants of East and Southeast Asia. The MIT Press, Cambridge,
USA, pp. 231.
Prashanth D, Asha MK, Amit A (2001). Antibacterial activity of Punica granatum.
Fitoterapia, 72:171–173.
Ridtitid W, Sae-wonga C,Wantana Reanmongkol W, MalineeWongnawaa M (2008).
Antinociceptive activity of the methanolic extract of Kaempferia galanga Linn. in
experimental animals. Journal of Ethnopharmacology, 118:225–230.
R´ıos JL, Recio MC (2005). Medicinal plants and antimicrobial activity, Journal of
Ethnopharmacology, 100:80-84.
Rout SD, Panda T, Mishra N (2009). Ethno-medicinal Plants Used to Cure Different Diseases
by Tribals of Mayurbhanj District of North Orissa. Ethno-Med, 3:27-32.
Roy S, Rao K, Bhuvaneswari, Giri A, Mangamoori LN (2010). Phytochemical analysis of
Andrographis paniculata extract and its antimicrobial activity. World J Microbiol Biotechnol,
26:85–91.
Ruslay S, Abas F, Shaari K, Zainal Z, Maulidiani, Sirat H, Israf DA, Lajis N (2007).
Characterization of the components present in active fractions of health gingers (Curcuma
xanthorrhiza and Zingiber zerumbet) by HPLC-DAD-ESIMS. Food Chemistry, 104:1183-
1191.
32
32
666
667
668
669
670
671
672
673
674
675
676
677
678
679
680
681
682
683
684
685
686
687
688
689
690
Russell AD (1991). Mechanisms of bacterial-resistance to non-antibiotics -food-additives and
food and pharmaceutical preservatives. Journal of Applied Bacteriology, 71:191–201.
Sader HS, Jones RN, Silva JB (2002). Skin and soft tissue infections in Latin American
Medical Centers: four year assessment of the pathogen frequency and antimicrobial
susceptibility patterns. Diagnostic Microbiology of Infectious Diseases, 45:281-288.
Sankaranarayanan S, Bama P, Ramachandran J, Kalaichelvan PT, Deccaraman M,
Vijayalakshimi M., Dhamotharan R., Dananjeyan B, Sathya Bama S (2010). Ethnobotanical
study of medicinal plants used by traditional users in Villupuram district of Tamil Nadu,
India. Journal of Medicinal Plants Research, 4:1089-1101.
Scot CN (2006). Morinda citrifolia (noni) Rubiaceae (coffee family) Species Profiles for
Pacific Island Agroforestry.
Shaari K, Safri S, Abas F, Lajis N, Israf DA (2004). A geranylacetophenone from the leaves
of Melicope ptelefolia Laboratory of Natural Products, Institute of Bioscience, Universiti
Putra Malaysia, Selangor, Malaysia
Shan B, Cai Y, Brooks JD, Corke H (2008). Antibacterial properties of polygonum
cuspidatum roots and their major bioactive constituents. Food chemistry, 109:530-537.
Shan B, Cai Y, Brooks JD, Corke H (2007). The in vitro antibacterial activity of dietary spice
and medicinal herb extracts, International Journal of Food Microbiology, 117:112-119.
33
33
691
692
693
694
695
696
697
698
699
700
701
702
703
704
705
706
707
708
709
710
711
712
713
714
715
Sibel R, (2003). Natural antimicrobials for the minimal processing of foods (1st
Eds),Woodhead Publishing Limited, 8:177-180.
Smid EJ, Gorris, LGM (1999). Natural antimicrobials for food preservation, In: Rahman, M.
S. (eds) Tropical Journal of pharmaceutical research, 5:557-560.
Snowdon JA, Buzby JC, Roberts T (2002). Epidemiology, Cost and Risk of Foodborne
Disease, in :Cliver, D.O., Riemann, H.P. (2nd Eds.), Foodborne Diseases. Academic Press.,
United Kingdom, pp. 45-46.
Sukrasno S, Fidriany I, Anggadiredja K, Handayani WA, Anam K (2011). Influence of drying
method on flavonoid content of Cosmos caudatus (kunth) leaves. Res. J. Med. Plant, 5:189-
195.
Tachibana Y, Kikuzaki H, Lajis N, Nakatani N (2001). Antioxidative activity of carbazoles
from Murraya koennigii leaves. Journal of Agriculture and Food Chemistry, 49:5589-5594.
Tang Y, Shi Y, Zhao W, Hao G, Le G (2008). Inhibition of food-borne pathogens by T1, a
novel antimicrobial peptide as a potential food preservative. International journal
food engineering, 4(4).
Taylor SL (2002). Disease Processes in Foodborne Illness, in :Cliver, D.O., Riemann, H.P.
(2nd Eds.), Foodborne Diseases. Academic Press., United Kingdom, pp. 11.
34
34
716
717
718
719
720
721
722
723
724
725
726
727
728
729
730
731
732
733
734
735
736
737
738
Tewtrakul S, Subhadhirasakul S, Puripattanavong J, Panphadung T (2003). HIV-1 protease
inhibitory substances from Boesenbergia pandurata Holtt. Songklanakarin. J. Sci.
Technology, 25:503–508.
Tewtrakul S, Yuenyongsawad S, Kummee S, Atsawajaruwan L (2005). Chemical components
and biological activities of volatile oil of Kaempferia galanga Linn. Songklanakarin Journal
of Science and Technology, 27:503–507.
Wattenberg, LW (1983). Inhibition of neoplasia by minor dietary constituents. Cancer Res.
(suppl.), 43:2448-2453
Webb AH, Tanner FW (1944). Effect of spices and flavoring material on growth of yeasts.
Food Research, 10:273-282.
Willey JM, Sherwood L, Woolverton CJ, Prescott LM (2008). Microbiology, (7th eds)
Prescott, Harley, and Klein's,pp 838-839.
Yadaz S, Vats V, Dhunnoo Y, Grover JK (2002). Hypoglycemic anti hyperglycaemic activity
of Murraya koennigii leaves in diabetic rats. Journal of Ethnopharmacology, 82:111-116.
Yasni S, Imaizumi K,Sin K,Sugano M,Nonaka G (1994). Identification of an active principle
in essential oils and hexane-soluble fractions of Curcuma xanthorrhiza Roxb. showing
triglyceridelowering action in rats. Food Chem. Toxicol, 32:273-278.
Zainol NA, Voo SC, Sarmidi MR, Aziz RA (2008). PROFILING OF CENTELLA ASIATICA
(L.) URBAN EXTRACT. The Malaysian Journal of Analytical Sciences, 12:322 -327.
35
35
739
740
741
742
743
744
745
746
747
748
749
750
751
752
753
754
755
756
757
758
759
760
761
762
763
Zurina H, Mun FY, Mariam A, Ahmad P M, Yusof (2010). Antidiabetic Properties and
Mechanism of Action of Gynura procumbens Water Extract in Streptozotocin-Induced
Diabetic Rats. Molecules, 15:9008-9023
36
36
764
765
766
767
768769770
771