Research Article (Open access) |
---|
SSR Inst. Int. J.
Life Sci., 6(1): 2449-2454,
January 2020
Antibacterial Activity of Water Extract From
Some Folk Medicine at Bidoup-Nuiba National Park, Lam
Dong, Vietnam
Pham Minh Nhut1,2*, Dang Thi Phuong
Thao1
1Department of Molecular
and Environmental Biotechnology, Faculty of Biology, University of Science,
Vietnam National University, Ho Chi Minh City, Vietnam
2Hutech Institute of
Applied Science, Ho Chi Minh University of Technology, Vietnam
*Address
for Correspondence: Dr. Pham Minh Nhut,
Department of Molecular and Environmental Biotechnology, Faculty of Biology,
University of Science, Vietnam National University Ho Chi Minh City, Vietnam227
Nguyen Van Cu Street, District 5, Ho Chi Minh City,
Vietnam
E-mail: dtpthao@hcmus.edu.vn
ABSTRACT-
Background: Folk
medicinal plants were used to treat some common diseases such as diarrhea,
wound healing by ethnic minorities at Bidoup-Nui Ba National Park, Lam Dong province, Vietnam. However, the
usage of medicinal plants was mainly based on experience and there was no
scientific basis to prove the biological activity of medicinal plants. This
study aims to evaluate the biological activity of some folk medicinal plants used by ethnic minorities.
Methods: In this
study, 9 samples of folk medicinal plants that ethnic minorities used to treat
diarrhea at Bidoup-Nui Ba
National Park, Lam Dong province were water extracted and evaluated the
antibacterial activity against 16 indicators bacterial sp. by well diffusion
agar. The water extracts were chemically
tested for the presence of different constituents, including carbohydrates,
alkaloids, saponins, cardiac glycosides, anthraquinone glycosides, flavonoids,
phenolic compounds, tannin, steroids and amino acids
by using standard methods.
Results: The results showed that water extract from Medinilla septentrionalis had the
highest antibacterial activity (against 16/16 indicator bacteria) with inhibition
diameter zone from 9.3 mm11.0 mm, 4/9 water extracts had weak activity and 4/9
extracts had no antibacterial activity. The preliminary phytochemical
screening of the water extracts showed the presence of flavonoids
and tannins.
Conclusion: Although the medicinal plants have been used widely
by ethnic people in the type of water extract,
this study showed that water
is not a good solvent for extracting biological compounds from the medicinal
plants. Further studies on theses plants should be carried out either for
evaluating their antibacterial activity or investigating their antidiarrheal activity.
Key Words: Antibacterial activity, Antidiarrhea, Bidoup-Nui Ba, Folk medicine, Water extract
INTRODUCTION- Vietnam is a tropical country with an abundant
and diverse medicinal plant. According to recent statistics, the flora of
Vietnam has over 10,000 species. Vietnam
has about 3,200 species of herbs. Although the sources of medicinal plants in
our country are abundant, the use and exploitation are limited, especially the
folk medicinal plants and indigenous knowledge. Folk medicinal plants are
widely used by the ethnic people in the treatment of diseases by cooked with
water until close to dry and then thoroughly drained for use [1-3].
Bidoup-Nui Ba National Park is a
large biodiversity center of Vietnam with 461 species of medicinal plants. K'Ho people, the main ethnic minority live here,
often use plants, leaves to treat some diseases such as diarrhea, ulcers,
abrasions, etc [4].
However, there have been few studies on folk medicinal plants at Bidoup-Nui Ba National.
Diarrhea is caused by many
reasons including bacterial infections such as Salmonella enteritidis, Salmonella typhi, Shigella flexneri, E. coli, Vibrio cholerae, Clostridium difficile
or chemicals such as castor oil or magnesium sulfate [5-7].
To treat diarrhea, some methods have been used such as oral rehydration therapy, antibiotics and gut
motility suppressing agents or pharmacological agents, etc. The usage is depending on the cause and the severity of
the disease [8-10]. However, medicine like loperamide,
diphenoxylate, racecadotril
might cause undesirable side effects as vomit, dry mouth, abdominal discomfort,
constipation and headache [11-13]. Recently, herbal remedies are great applications of interest. Most of medicinal plants have alkaloid, steroid, tannin and
phenol which are stored in some different part of the plants [14].
These metabolites are well known as
antibacterial substances, which prevent bacterial
infection into the plants. Although the antibacterial mechanism still was not
fully reported, it is known that the metabolite inhibit bacterial DNA/RNA
synthesis, interfere bacterial metabolism, and destroy bacterial cell membrane [15,16]. In this study, we evaluated the
antimicrobial activity of the folk medicinal plants against some bacteria that cause diarrhea.
MATERIALS AND METHODS
Folk
medicinal plants and extraction- Medicinal plants (Elephantopus mollis Kunth, Lantana camara L., Euodia lepta (Spreng.) Merr., Acorus tatarinowii Schott., Calamus sp., M. septentrionalis (W.W.Sm) M.P. Nayar., Polygala paniculata L., Podocarpus imbricatus Blume., Eupatorium odoratum L.) (Table 1) were collected from Bidoup-NuiBa
National Park, Lam Dong province, Vietnam and identified at Department of
Ecology and Evolutionary Biology, Faculty of Biology, University of Science,
VNU-HCMC from August 2016 to September 2017. These samples were extracted according to Akinyemi et al. [4]
with modification. Samples were dried, powdered and extracted
with water by immersion method [1:10 (w/v)] at room temperature for 4 hours. The extracts were filtered and
evaporated at 400C using a rotary evaporator to constant weight. The
extracts were then dissolved in sterile distilled water and stored in a
sterilized bottle in 40C temp.
Table 1: Medicinal plants from Bidoup-Nui
Ba National Park, Lam Dong province, Vietnam
Plants name |
Vietnamese name |
Used plant parts |
Elephantopus
mollis Kunth |
Chan voi mem |
Bud |
Lantana camara L. |
Bong oi |
Bud |
Euodia
lepta (Spreng.) Merr. |
Ba chạc |
Bud and leaf |
Acorus
tatarinowii Schott |
Thachxuongbo |
Base |
Calamus sp. |
May rung |
Bud |
Medinilla septentrionalis (W.W.Sm) M.P.Nayar |
Xidranguom |
Bud and leaf |
Polygala paniculata L. |
Kichnhuthom |
Whole tree |
Podocarpus
imbricatus Blume |
Bach tung |
Stem core, leaf |
Eupatorium odoratum L. |
Co hoi |
Root |
Indicator bacterial species- Indicator bacteria were provided by the
Department of Molecular and Environmental Biotechnology, Faculty of Biology,
University of Science, VNU including 3 strains of Shigella sp. (S. sonnei, S. flexneri, and
S. boydii), 4 strains of Salmonella sp. (S. typhimurium, S. typhii,
S. dublin, S. enteritidis),
3 strains of Escherichia coli (Enterotoxigenic E. coli (ETEC), E. coli O157:H7,
E. coli), 3
strains of Vibrio sp. (V. cholerae,
V. parahaemolyticus), 2 strains of Listeria sp. (L.
monocytogenes, L. innocua)
and Staphylococcus aureus.
Antibacterial activity- Antibacterial activity of 9 water extracts
was evaluated by well diffusion agar method [17,18]. Indicator bacterial sp. was enriched in the suitable medium at
temperature 370C at 24 hours. 100μl of diluted bacteria (106 cfu/ml1) solution was spread on agar
medium until drying. Then, wells (6 mm diameter) were made in each plate by
using sterile metal cylinders. 100 μl of every
water extract (100 mg/ml1) were dropped into the wells. Sterile distilled water was used as a
control. Plates were incubated at 37.0±0.10C at 24 hours. The
diameter of the inhibition zone (mm) was measured. Each experiment was triplicated and collected data were subjected to
statistical analysis.
Preliminary phytochemical
analysis- Total 9 water
extracts were identified [17] the presence of constituents, including
carbohydrate (Molisch, Fehling and Barfoed test), alkaloid (Mayer, Dragendorff,
Hager and Wagner test), saponin (foam test), cardiac
glycoside (Legal and Keller-Killiani test), anthraquinone glycoside (Bontrager
test), flavonoid (alkaline and ferric chloride test),
tannin (ferric chloride and lead acetate test), steroid (Salkowski
and Libermann Burchard
test), amino acid (ninhydrin test).
Statistical Analysis- Statistical analysis values were expressed as
mean±standard deviation. Mean values were evaluated
by Analysis of Variance. Duncan test was used to determine the statistical
significance (P< 0.05).
RESULTS
Identification of the extractive efficiency
of water extracts- Water
is a solvent that used to extract the compound from medicinal plants by KHo people at Bidoup-Nui Ba National Park. We, therefore utilized it for preparation of the plant
extracts. The extractive
efficiency of 9 medicinal plants with water was a statistically significant
difference (p <0.05) (Table 2). The extraction efficiency of E. lepta was highest (29.52%), while the
extraction efficiency from E. mollis, P. imbricatus, L. camara was low (<20%).
Table 2: The
extractive efficiency of water extract
Medicinal
plants |
Extractive
efficiency of water extract (%) |
L. camara |
12.36±0.43ef |
A. tatarinowii |
25.74±0.52ab |
E. lepta |
29.52±1.04a |
Calamus sp. |
21.69±0.44d |
M. septentrionalis |
20.18±1.19de |
P. paniculata |
20.53±1.09d |
E. mollis |
15.15±0.46ef |
P. imbricatus |
11.99±0.43f |
E. odoratum |
25.46±2.04bc |
Antibacterial activity of water extracts- The results of the antimicrobial activity of
water extracts against indicator bacteria showed that 4/9 plant extracts did
not inhibit indicator bacteria, while another 5/9 water extracts showed
different antibacterial activity (Table 3). In particular, M. septentrionalis water extract showed the strongest antibacterial
activity (inhibition to 16/16 indicator bacteria), P. paniculata, P. imbricatus, E. odoratum,
and E. mollis
water extract inhibited 6/16, 2/16, 1/16 and 1/16 indicator bacteria at 100
mg/ml concentration, respectively.
Table 3: Antibacterial inhibition zone of water extracts against
indicator bacteria (mm)
Bacterial sp. |
LC |
AT |
EL |
CA |
MS |
PP |
EM |
PI |
EO |
E. coli O157:H7 |
NA |
NA |
NA |
NA |
11.00±0.50 |
NA |
NA |
NA |
NA |
E. coli |
NA |
NA |
NA |
NA |
10.17±1.15 |
8.17±0.29 |
NA |
NA |
NA |
ETEC |
NA |
NA |
NA |
NA |
9.83±0.29 |
8.83±0.39 |
NA |
NA |
NA |
L. innocua |
NA |
NA |
NA |
NA |
10.17±0.58 |
NA |
NA |
NA |
NA |
L. monocytogenes |
NA |
NA |
NA |
NA |
12.83± 0.29 |
10.00±0.00 |
NA |
NA |
NA |
S. dublin |
NA |
NA |
NA |
NA |
11.00±0.50 |
NA |
NA |
NA |
NA |
S. enteritidis |
NA |
NA |
NA |
NA |
10.75±0.35 |
NA |
NA |
NA |
NA |
S. typhii |
NA |
NA |
NA |
NA |
9.67±0.29 |
9.00 ± 1.00 |
NA |
9.83±0.29 |
NA |
S. typhimurium |
NA |
NA |
NA |
NA |
10.83±0.58 |
NA |
NA |
NA |
NA |
S. boydii |
NA |
NA |
NA |
NA |
11.75±0.40 |
NA |
NA |
NA |
13.17±0.29 |
S. flexneri |
NA |
NA |
NA |
NA |
9.83±0.29 |
NA |
12.17±0.29 |
NA |
NA |
S. sonnei |
NA |
NA |
NA |
NA |
9.33±0.60 |
NA |
NA |
11.5±0.00 |
NA |
V. cholerae |
NA |
NA |
NA |
NA |
10.00±1.00 |
NA |
NA |
NA |
NA |
V. parahaemolyticus |
NA |
NA |
NA |
NA |
9.83±0.76 |
8.33±0.59 |
NA |
NA |
NA |
S. aureus |
NA |
NA |
NA |
NA |
9.83±0.29 |
NA |
NA |
10.00±0.00 |
9.5±0.00 |
E. feacalis |
NA |
NA |
NA |
NA |
9.50±0.50 |
11.50±1.50 |
NA |
NA |
NA |
LC: Lantana camara; MS:
Medinilla septentrionalis; PP: Polygala paniculata;
AT: Acorustatarinowii; EM: Elephantopusmollis; PI:
Podocarpusimbricatus; EL: Euodialepta; EO: Eupatorium odoratum; CA: Calamus sp.
Phytochemical
identification of the water extracts- The results of the chemical composition of water extracts from 9
medicinal plants showed that carbohydrates presented at all water extracts. In
addition, there was the presence of several biologically active compounds,
including alkaloids, tannins, and flavonoids (Table
4).
Table
4: Phytochemical constituents of water extracts
Constituents |
Test |
LC |
AT |
EL |
CA |
MS |
PP |
EM |
PI |
EO |
Carbohydrate |
Molisch |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
Fehling |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
|
Barfoed |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
|
Alkaloid |
Mayer |
+ |
|
+ |
|
|
+ |
|
|
|
Dragendorff |
+ |
|
+ |
|
|
+ |
|
|
|
|
Hager |
+ |
|
+ |
|
|
+ |
|
|
|
|
Wagner |
+ |
|
+ |
|
|
+ |
|
|
|
|
Saponin |
Foam |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
Cardiace
glycoside |
Legal |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
Keller-Killiani |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
|
Anthraquinone
glycoside |
Borntrager |
|
|
|
|
+ |
+ |
+ |
+ |
+ |
Flavonoid |
Alkaline |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
Ferric chloride |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
|
Tannin |
Ferric chloride |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
Lead acetate |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
|
Steroid |
Salkowski |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
Libermann-Burchard |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
|
Amino acid |
Ninhydrin |
|
|
|
|
|
|
|
|
|
(+): positive; (): negative
LC: Lantana
camara; MS:
Medinilla septentrionalis; PP:
Polygala paniculata;
AT: Acorustatarinowii;
EM: Elephantopusmollis; PI: Podocarpusimbricatus; EL: Euodialepta; EO: Eupatorium odoratum;
CA: Calamus
sp.
DISCUSSION- Understanding the effectiveness of traditional plants, which used by ethnic people are important in order to sustainable mining
folk medicinal plants and develop the usage, this study aimed to evaluate the antibacterial effectiveness of folk
medicinal water extract that used by the K'Ho ethnic
group in the treatment of some common diseases including diarrhea.
To elucidate the
antibacterial activity of the plants and the indicator bacterial sp. belonged
to the gastroenterogenic bacteria including S. typhii caused typhoid; S. enteritidis, S. flexneri
and E. coli caused diarrhea; S. sonnei, S. boydii caused dysentery; Vibrio sp. caused many dangerous
intestinal diseases such as cholera (V. cholerae) [17,18]. The results showed that the antimicrobial activity of water extracts
from 9 folk medicinal plants was very low, except M. Septentrionalis water extract had high antibacterial activity.
This study showed
that water was not a good solvent to extract antimicrobial compounds for those
plants in this study. However, in the daily file, ethnic people, mostly boiled the medicinal
plants in water and used to treat diarrhea. Our study results suggested that
the usage even based on people experiences but may not high efficiency in
treatment as the antibacterial activity is low in 8/9 plants except M. septentrionalis.
Besides,
our results of
the preliminary phytochemical
screening also provided the scientific evidences that the plants
contain some well known antibacterial
compounds such as flavonoid phenolic compound. The plants also contain some compounds that can precipitate protein, which functions on the electrolyte and reduce small intestine transit and intestinal
secretion such as tannin and flavonoid [19,20]. Taken together, the study suggested that the folk medicinal plants
play antidiarrheal activity via their antibacterial potential and their metabolite activity on electrolyte,
intestine. However, the traditional usages
should be improved by using different solvent for plant extracts.
CONCLUSIONS: In 9 folk medicinal plant samples at Bidoup-Nui
Ba National Park, only 5/9 water extracts had antibacterial
activity against diarrhea relating indicator bacteria and M. septentrionalis water extract had the strongest antibacterial
activity (19/19 indicator bacteria) at 100 mg/ml concentration.
This result showed that water is not a good solvent for
extracting biological compounds from all medicinal plants; therefore, it is
important to investigate before using water to extract from medicinal plants.
ACKNOWLEGDEMENT- We thank Vietnam
National University in Ho Chi Minh City for funding this study. We thank Directors of Bidoup-Nui
Ba National Park, Lam Dong Province for facilitating
to collect samples. We thank BSc. Pham Thi Thao, Lam Pham Hue Tam at Ho Chi Minh City University of
Technology has supported the experiment.
CONTRIBUTION
OF AUTHORS
Research concept- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Research design- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Supervision- Dr. Dang Thi
Phuong Thao
Materials- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Data collection- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Data analysis and
Interpretation- Dr.
Dang Thi Phuong Thao, Pham
Minh Nhut
Literature search- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Writing article- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Critical review- Dr. Dang Thi
Phuong Thao
Article editing- Dr. Dang Thi
Phuong Thao, Pham Minh Nhut
Final approval- Dr. Dang Thi
Phuong Thao
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