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Correspondence:
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Contents lists available at Article URL
KKU Journal of Basic and Applied Sciences Journal homepage: jbas.kku.edu.sa
Vol 3, 2 (2017) 31-37
P a g e | 31
Antibacterial and Molluscicidal Activities of Euphorbia spp.
Grown Naturally in Aseer Region, Saudi Arabia
Mahmoud Moustafaa,b*, Dema Al-Shahranic, Osama Mostafaa,d and Sulaiman Alrummana aDepartment of Biology, Faculty of Science, King Khalid University, Abha, Saudi Arabia
bDepartment of Botany, Faculty of Science, South Valley University, Qena, Egypt cDepartment of Biology, Faculty of Science and home economics, Bisha University, Saudi Arabia dDepartment of Zoology, Faculty of Science, Ain Shamis University, Egypt
Keywords: B. arabica, E. peplus, E. prostrata, E. terracina.
1. INTRODUCTION
Genus Euphorbia is one of the biggest groups in the flowering
plants comprising the third largest genus and has many forms
including herbs, shrubs and trees ranging from succulent to
non-succulent plants [1]. The recorded number of species in
the genus of Euphorbia was about 2,160 that had been
classified in the order Malpighiales and in family
Euphorbiaceae. Euphorbs plant is found in many area across
the globe including the tropical, subtropical localities of Africa
and in the Americas and in temperate zones. Euphorbia species
have their own economic importance so why the Euphorbs
contribute to the floristic wealth of tropical and subtropical
countries and for many other countries all over the world
whereas the family composed of 322 genera and 8910 species
[2]. Recently, many of Euphorbs plant have been used in
medicine plants, for example, the stem is used for healing
bronchitis, asthma, and various other lung problems and
showed anti-pyretic, analgesic, sedative, anxiolytic, inhibitory
effect on platelet aggregation, anti-inflammatory behavior,
antioxidant, and
antimutagenicity [3], [4]. The way for the elimination of
human pathogenic schistosomes in the country of Saudi Arabia
was mainly depend on regular chemotherapy of infected
people and snail management by using synthetic agents
molluscicides [5]. However, it was reported that the synthetic
agents molluscicides had many disadvantages [6], [7]. On the
other hand, the antibacterial effect of some Euphorbia spp. had
been studied previously, such as solvents extract obtained from
Euphorbia hirta and Euphorbia heterophylla against a range
of clinically resistant pathogens by using broth microdilution
method and found that it could be used as natural antibacterial
agents [8], [9]. The traditional uses of many herbal plants
usually not validated scientifically so scientific methodology
could provide valuable leads to discover new drugs and
expanded utilization of them in the future. It was found that
about 119 of identified chemical compounds obtained from 91
species are mainly used in drugs manufacture in many
countries [10]. Combating the multidrug resistant
microorganism such as Staphylococcus, S. dysenteriae, S.
enteritidis, S. paratyphi etc using natural product are the main
Received: 30 November 2017/ Revised: 27 December 2017/ Accepted: 09 January 2018/ Published: 01 July 2018
Abstract: An in vitro antibacterial activities of some solvents extracts gained from Euphorbia prostrata, Euphorbia
peplus and Euphorbia terracina were tested against numbers of human pathogenic bacteria using well diffusion method.
The anti-molluscicidal activity against Biomphalaria arabica was determined using various concentration of aqueous
extract from Euphorbia spp. The result of antibacterial inhibition activities revealed that five solvents extracts had
various degrees of antibacterial activities. The methanol and petroleum ether extracts gained from Euphorbia prostrata
had a greater effect on Pseudomonas aeruginosa than Euphorbia peplus and Euphorbia terracina. Ethanol extract from
Euphorbia peplus, methanol extract from Euphorbia prostrata and ethanol extract from Euphorbia terracina strongly
ceased the growth of Klebsiella pneumoniae and Shigella sp. The molluscicidal activity results showed that the lowest
concentration used (0.5%) had the earliest 100% mortality after 6 hours for Euphorbia prostrata extract and the latest
100% mortality for the same concentration was noted after 48 hours of exposure for Euphorbia terracina. For the
highest concentration tested (3%), 100% mortality was occurred after only 1 hour of exposure to Euphorbia prostrata,
Euphorbia peplus and Euphorbia terracina extracts. The present study clearly indicated that Euphorbia spp. could be
used as an alternative promising natural antibacterial and molluscicide agents.
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KKU Journal of Basic and Applied Sciences
P a g e | 32
goals of many researchers all over the world [11], [12], [13].
Aseer region, KSA, characterized by unique environmental
condition and the landscape has great variety of Euphorbia
plants and many of them have not been examined yet for
various aspects of biology. Consequently, this research
focuses on the antibacterial and antischistosomal activities of
three species of Euphorbia including Euphorbia prostrata
Aiton, Euphorbia peplus L. and Euphorbia terracina L.
growing naturally in Aseer region, KSA as a first report.
2. MATERIALS AND METHODS
Plant materials
Three species of healthy plants from species Euphorbia
namely E. prostrata, E. peplus and E. terracina were collected
from two different locations, E. terracina from El Sauda
Mountains (2270 meters height) and E. prostrata and E. peplus
from the garden of King Khalid University (2125 meters
height).The specimens were identified and the voucher kept in
the herbarium of Biology department, Faculty of Science,
King Khalid University.
Preparation of leaves extracts from the three Euphorbia spp.
for antibacterial activity
45 gms from leaves of E. prostrata, E. peplus and E.terracina
were washed using distilled water and subjected to the drying
in shade for 15 days and the dried plant materials were crushed
into fine particles (powder) using a mixer [14]. Extraction was
accomplished by adding 20 ml from acetone, chloroform,
ethanol, methanol and petroleum ether to the half-gram (0.5 g)
of air-dried powdered leaves materials. Samples were kept in
rotary shaker at 99 rpm at room temperature for 48 hours. The
resultant extract was filtered using Whatman filter paper
(No.1) and placed in incubator at 49 C until the solvent was
evaporated completely. Each extract was weighed, dissolved
in sterile dimethyl sulfoxide (DMSO) and subjected to the
antibacterial activity test [15].
Test organisms used
Five pathogenic organisms including Klebsiella pneumoniae,
Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus
aureus and Shigella sp. are used in this study. All bacterial
strains were first sub-cultured in nutrient broth at 37 C for 24
hours. All strains were obtained from the Microbiology
Laboratory, Faculty of Science and Microbiology Laboratory,
Faculty of Medicine, King Khalid University, Kingdom of
Saudi Arabia.
Screening for antibacterial activity
The efficacy of antibacterial activities of various solvent
extract including acetone, chloroform, ethanol, methanol and
petroleum ether gained from dried leaves of E. prostrata, E.
peplus and E. terracina and tested against K. pneumoniae, P.
mirabilis, P. aeruginosa, S. aureus and Shigella sp. were
accomplished by using the well diffusion method [16]. 20 ml
from sterilized Mueller Hinton agar (Oxoid, England) media
was poured into sterile Petri dish and 0.1 ml of standardized
inoculums of each test bacterium swapped onto agar. 6 mm
diameter well was cut from the agar and each well filled with
0.1 ml of the plant extract. Each extract was assayed in
triplicate and sterile and 0.1 ml from dimethyl sulfoxide
(DMSO) used as negative control and cefoxitin (30 mcg) as
positive controls. All Petri dishes were incubated at 30 C for
24 h and the antibacterial activity was measured by measuring
the size of the inhibition zones [17].
Statistical analysis
One-way analysis of variance (ANOVA) was applied to
determine statistically significant differences amongst tested
extracts. A p ≤ 0.05 was considered statistically significant.
Preparation of plant extracts for molluscicidal activity
0.5, 1, 1.5, 2, 2.5, and 3 grams from air-dried leaves of E.
prostrata, E. peplus and E. terracina were ground into fine
particles and each of them was added to 100 ml of water at
room temperature for 72 hours with shaking. The extracts were
filtered and used for the study of molluscicidal activity.
Snails Collection
Adults freshwater snails Biomphalaria arabica (5-8 mm in
shell diameter) were collected from freshwater bodies in Besha
region, Saudi Arabia and were kept in plastic containers each
contain one liter of dechlorinated water for at least three days
for acclimatization under laboratory conditions. The water was
changed every day and the snails were fed daily on fresh
lettuce leaves.
Molluscicidal activity
Tests were performed in 24-well plates whereas the snails were
placed individually in the wells with about two ml of the tested
concentration. To avoid the snails to be out of the wells, the
wells were covered and the gap between the liquid and the
cover didn't permit the snails to leave. Six groups of snails
were exposed to concentrations of 0.5, 1, 1.5, 2, 2.5, and 3 %
of E. prostrata, E. peplus and E. terracina extracts for 1, 2, 3,
6, 12, 24 and 72 hours, then the snails were transferred to
dechlorinated water for another 24 hours for recovery. After
the recovery period, the snails were examined with the aid of
stereoscopic microscope to calculate the living and nonliving
snails to measure the rate of mortality. The deaths of the snails
were confirmed by observation of contraction of the soft parts
within the shell, absence of muscle contractions in response to
needle probe and change in the shell color. Control
experiments were performed with dechlorinated water [18].
3. RESULTS AND DISCUSSION
Antibacterial properties of Euphorbia spp. extracts against
P. mirabilis
The in vitro antibacterial activities of acetone, chloroform,
ethanol, methanol and petroleum ether of dried leaves of E.
prostrata, E. peplus and E. terracina extracts against P.
mirabilis was evaluated using well diffusion methods.
According to the results of the study (Table I), all solvents
extract significantly inhibited the growth of P. mirabilis.
Among the 3 Euphorbia spp. tested, the acetone and ethanol
leaf extracts of E. peplus showed the highest antibacterial
activity with a zone of inhibition 24.67±2.91 mm at 0.37 g/ml
concentration. The chloroform, methanol and petroleum ether
extract of the dried leaves of E. prostrata had a slightly lower
antibacterial activity than the acetone and ethanol leaf extracts
of E. peplus with a zone of inhibition ranging from 24.33±0.67
to 24.00±1.00 mm. Similarly, ethanol and petroleum ether
extract from E. terracina had also a slightly lower antibacterial
activity with a zone of inhibition 24.33±2.33 mm and
24.33±0.66 mm respectively. On the other hand, the acetone
and ethanol extract of E. prostrata and acetone, chloroform
and methanol extract had a moderate antibacterial activity with
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KKU Journal of Basic and Applied Sciences
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a zone of inhibition ranging from 23.33±1.67 mm to
20.00±0.00 mm.
TABLE I: SUSCEPTIBILITY OF P. MIRABILIS
Mean diameter of zone of inhibition (mm) ± SE at concentration of 0.37 g/ml (95% Confidence
interval for mean; lower - upper bound)
E. peplus E. prostrata E. terracina P.C N.C
24.67±2.91
(12.16-37.17)
23.33±1.67
(16.16-30.50)
20.00±0.00*
(20.00-20.00)
27.00±0.58
(24.52-29.49)
NI
19.33±2.96**
(6.59-32.08)
24.33±0.67
(21.46-27.20)
22.33±1.45
(16.08-28.58)
27.00±0.58
(24.52-29.49)
NI
24.67±2.40
(14.32-35.01)
20.67±0.67*
(17.70-23.53)
24.33±2.33
(14.29-34.37)
27.00±0.58
(24.52-29.49)
NI
23.00±2.65
(11.62-34.38)
24.00±1.00
(19.60-28.30)
21.67±1.67*
(14.40-28.84)
27.00±0.58*
(24.52-29.49)
NI
24.33±2.33
(14.29-34.37)
24.33±1.33
(18.50-30.07)
24.33±0.66
(21.46-27.20)
27.00±0.58
(24.52-29.49)
NI
P.C. (positive control, Cefoxitin-30 mcg); N.C. (negative control, DMSO, dimethyl sulfoxide)
NI = No inhibition; * p<0.05, ** p<0.01, represent significant difference compared with positive control.
However, chloroform extract from E. peplus showed lowest
antibacterial effect with their respective diameters of
inhibition zones of 19.33±2.96 mm. No antibacterial activity
was observed against any bacterial for negative control.
Antibacterial properties of Euphorbia spp. extracts against
P.aeruginosa In Table (II), chloroform, petroleum ether, methanol, ethanol
and acetone extracts of dried leaves of E. prostrata, E. peplus
and E. terracina showed varied antibacterial activity against
P. aeruginosa with zones of inhibition ranging from
17.67±2.33 mm to 26.33±1.33mm. Methanol extracts from E.
prostrata showed the highest antibacterial activities with a
zone of inhibition 26.67±.88 mm while acetone from E. peplus
showed modest activity with a zones of inhibition 23.00±2.89
mm. Ethanol and methanol
extracts from E. peplus, acetone extract from E. prostrata and
acetone and petroleum ether extracts from E. terracina showed
a slightly lower antibacterial activity with a zone of inhibition
ranging from 21.00±1.00 mm to 20.00±0.00 mm. Chloroform
extracts from E. peplus showed lowest activity with zones of
inhibition 17.67±2.33 mm. No antibacterial activity was
observed against any bacterial for negative control.
TABLE II: SUSCEPTIBILITY OF P. AERUGINOSA
Mean diameter of zone of inhibition (mm) ± SE at concentration of 0.37 g/ml (95%
Confidence interval for mean; lower - upper bound)
Solvent
stra
ins
N.C
P.C E. terracina E. prostrata E. peplus
NI 20.33±0.67
(17.46-23.20)
20.00±0.00
(20.00-20.00)
21.66±1.66
(14.40-28.84)
23.00±2.89
(10.58-35.42)
Acetone
P.
aer
ug
ino
sa
NI 20.33±0.67
(17.46-23.20)
23.33±1.67
(16.16-30.50)
23.33±1.66
(16.16-30.50)
17.67±2.33
(7.63-27.71)
Chloroform
NI 20.33±0.67
(17.46-23.20)
23.333±1.67
(16.16-30.50)
20.00±0.00**
(2.00-20.00)
21.33±1.33
(15.50-27.07)
Ethanol
NI 20.33±0.67
(17.46-23.20)
23.33±1.67
(16.16-30.50)
26.67±0.88*
(22.87-30.46)
21.00±1.00
(16.60-25.30)
Methanol
NI 20.33±0.67
(17.46-23.20)
21.33±1.33*
(15.50-27.07)
26.33±1.33
(20.50-32.07)
19.67±2.60
(8.47-30.87)
Petroleum ether
P.C. (positive control, Cefoxitin-30 mcg); N.C. (negative control, DMSO, dimethyl sulfoxide)
NI = No inhibition; * p<0.05, ** p<0.01, represent significant difference compared with positive control
.
Antibacterial properties of Euphorbia spp. extracts against
S. aureus
Table (III) showed that the results of solvents extracts of three
Euhorbia spp. against S. aureus. It inhibited by the plant
solvent extract between 24.67±1.76 mm to 20.00±0.00 mm.
Ethanol extracts from E. terracina had the highest antibacterial
activities against staphylococcus with zones of inhibition of
24.67±1.76 mm, followed by those prepared in acetone and
methanol extract form E. terracina and E.
peplus with a zone of inhibition between 24.33±2.33 and
24.00±0.67 mm. Acetone and petroleum ether extract from E.
prostrata and E. terracina showed a slightly lower antibacterial
activity with a zone of inhibition ranging 23.33±1.67 mm to
23.00±1.53 mm. Chloroform and petroleum ether extracts
from E. peplus showed lowest activity with zones of inhibition
20.00±0.00 m
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TABLE III: SUSCEPTIBILITY OF S. AUREUS
Mean diameter of zone of inhibition (mm) ± SE at concentration of 0.37 g/ml (95%
Confidence interval for mean; lower - upper bound)
Solvent stra
ins
N.C
P.C E. terracina E. prostrata E. peplus
NI 28.00±0.58
(25.52-30.48)
23.00±1.53
(16.43-29.57)
23.33±1.67*
(16.16-30.50)
24.00±2.65**
(12.62-35.38)
Acetone
S.
au
reu
s
NI 28.00±0.58
(25.52-30.48)
23.33±1.67
(16.16-30.50)
22.67±2.67*
(11.19-34.14)
20.00±0.00*
(20.00-20.00)
Chloroform
NI 28.00±0.58
(25.52-30.48)
24.67±1.76
(17.08-32.26)
21.67±1.76*
(14.08-29.26)
22.00±2.52
(11.17-32.83)
Ethanol
NI 28.00±0.58
(25.52-30.48)
24.00±0.67
(21.46-27.20)
23.00±1.53
(16.43-29.57)
24.33±2.33**
(14.29-34.37)
Methanol
NI 28.00±0.58
(25.52-30.48)
23.33±1.67
(16.16-30,50)
23.33±1.67*
(16.16-30.50)
20.00±0.00
(20.00-20.00)
Petroleum ether
P.C. (positive control, Cefoxitin-30 mcg); N.C. (negative control, DMSO, dimethyl sulfoxide)
NI = No inhibition; * p<0.05, ** p<0.01, represent significant difference compared with positive control.
Antibacterial properties of Euphorbia spp. extracts
against Shigella sp.
Table (IV) showed that all solvents extracts gained from dried
leaves of E. peplus, E. peplus and E. terracina had varied
antibacterial activity against Shigella sp. in the range between
25.67±1.20 mm to 20.00±0.00 mm. Methanol extract from E.
peplus and ethanol extract from E. terracina had the highest
antibacterial activities with inhibition zone of 25.67±1.20 mm
and 25.00±0.00 mm
respectively. Acetone, methanol and petroleum ether extract
from E. prostrata showed a slightly lower antibacterial activity
with a zone of inhibition between 24.67±0.33 mm to
23.00±1.53 mm. Acetone, methanol and petroleum ether
extracts from E. terracina and chloroform, ethanol and
petroleum ether extracts from E. peplus showed lowest activity
with zones of inhibition ranging from 20.67±2.96 to
20.00±0.00 mm. No antibacterial activity was observed
against any bacterial for negative control.
TABLE IV: SUSCEPTIBILITY OF SHIGELLA SP.
Mean diameter of zone of inhibition (mm) ± SE at concentration of 0.37 g/ml (95%
Confidence interval for mean; lower - upper bound)
Solvent stra
ins
N.C
P.C E. terracina E. prostrata E. peplus
NI 29.00±0.58
(26.52-31.48)
20.00±0.00*
(20.00-20.00)
23.00±1.53**
(16.43-29.57)
21.33±1.86**
(13.35-29.32)
Acetone
Sh
igel
la s
p.
NI 29.00±0.58
(26.52-31.48)
20.00±0.00*
(20.00-20.00)
22.67±1.45**
(16.42-28.92)
20.67±1.20**
(15.40-25.84)
Chloroform
NI 29.00±0.58
(26.52-31.48)
25.00±0.00**
(25.00-25.00)
23.00±2.00*
(14.39-31.61)
20.67±2.96*
(7.92-33.41)
Ethanol
NI 29.00±0.58
(26.52-31.48)
23.33±1.67**
(16.16-30.50)
24.00±2.08**
(15.04-32.96)
25.67±1.20**
(20.40-30.84)
Methanol
NI 29.00±0.58
(26.52-31.48)
21.67±1.67
(14.40-28.84)
24.67±0.33**
(23.23-26.10)
20.00±0.00**
(20.00-20.00)
Petroleum ether
P.C. (positive control, Cefoxitin-30 mcg); N.C. (negative control, DMSO, dimethyl sulfoxide)
NI = No inhibition; * p<0.05, ** p<0.01, represent significant difference compared with positive control.
Antibacterial properties of Euphorbia spp. extracts
against K. pneumonia
E. peplus, E. peplus and E. terracina solvents extracts against
K. pneumoniae had prominent antibacterial activity with
inhibition zones ranging between 25.67±1.45 mm to
21.00±3.51 mm as shown in (Table V). Ethanol extract from
E. peplus and methanol extract from E. prostrata had the
highest antibacterial activities with zones of inhibition
25.67±1.45 mm and 25.00±0.00 mm respectively. Acetone
and methanol extracts from E. peplus, ethanol and petroleum
ether extracts from E. prostrata and chloroform, methanol and
petroleum ether extracts from E. terracina showed slightly less
activity with zones of inhibition ranging from 23.33±1.67 to
23.00±1.53 mm. Chloroform extract from E. prostrata showed
the lowest activity with zones of inhibition 20.00±0.00mm.
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TABLE V: SUSCEPTIBILITY OF K. PNEUMONIAE
Mean diameter of zone of inhibition (mm) ± SE at concentration of 0.37 g/ml (95%
Confidence interval for mean; lower - upper bound)
Solvent stra
ins
N.C
P.C E. terracina E. prostrata E. peplus
NI 29.33±0.33
(27.80-30.77)
21.67±1.67*
(14.40-28.84)
21.67±1.67**
(14.40-28.84)
23.33±1.67**
(16.16-30.50)
Acetone
K.
pn
eum
on
iae NI 29.33±0.33
(27.80-30.77)
23.33±1.67**
(16.16-30.50)
20.00±0.00*
(20.00-20.00)
21.00±3.51
(5.89-36.11)
Chloroform
NI 29.33±0.33
(27.80-30.77)
21.67±1.67*
(14.40-28.84)
23.00±1.53
(16.43-29.57)
25.67±1.45*
(19.42-31.92)
Ethanol
NI 29.33±0.33
(27.80-30.77)
23.33±1.67*
(16.16-30.50)
25.00±0.00*
(25.00-25.00)
23.33±3.28**
(9.21-37.46)
Methanol
NI 29.33±0.33
(27.80-30.77)
23.33±1.67*
(16.16-30.50)
23.00±1.53**
(16.43-29.57)
21.67±1.67**
(14.40-28.84)
Petroleum
ether
P.C. (positive control, Cefoxitin-30 mcg); N.C. (negative control, DMSO, dimethyl sulfoxide)
NI = No inhibition; * p<0.05, ** p<0.01, represent significant difference compared with positive control.
Antischistosomal activity
The present data revealed that aqueous leaves extract of E.
peplus, E. prostrata and E. terracina had a molluscicidal
activity against B. arabica snails, the intermediate host of S.
mansoni in Saudi Arabia (Table VI, VII, VIII). For E. peplus
extract, 100% mortality occurred after only 2 hours of snails'
exposure to concentrations 2, 2.5 and 3%. For 1.5%
concentration, 100% mortality appeared after 3 hours of
exposure. 100% mortality delayed to 24 hours for
concentrations 0.5 and 1% concentrations of the same extract.
Concerning the extract of E. prostrata, 100% mortality
appeared after 1, 2, 3 hours of exposure to extract
concentrations 3, 2.5 and 2%, respectively. After 6 hours
exposure to concentrations 0.5, 1, 1.5 % all snails were died.
The extract of E. terracina achieved 100% mortality at 1, 2, 6
and 48 hours of exposure to concentrations 3, 2.5, 2, 0.5%
respectively. For 1 and 1.5% concentrations, 100% mortality
was achieved after 24 hours of exposure.
TABLE VI: MORTALITY RATE (%) OF
B. ARABICA SNAILS EXPOSED TO DIFFERENT
CONCENTRATIONS OF E. PEPLUS EXTRACT
72
hours
48
hours
24
hours
12
hours
6
hours
3
hours
2
hours
1
hour
Conc./
time
100 0.00 0.00 0.00 0.00 0.00 0.5%
100 20.0 20.0 20 0.00 0.00 1%
100 20.0 0.00 1.5%
100 40.0 2%
100 60.0 2.5%
100 100 3% TABLE VII: MORTALITY RATE (%) OF
B. ARABICA SNAILS EXPOSED TO DIFFERENT
CONCENTRATIONS OF E. PROSTRATA EXTRACT
72
hours
48
hours
24
hours
12
hours
6
hours
3
hours
2
hours
1
hour
Conc./
time
100 0.00 0.00 0.00 0.5%
100 20.0 20.0 0.00 1%
100 40.0 20.0 0.00 1.5%
100 40.0 40.0 2%
100 60.0 2.5%
100 3% TABLE VIII: MORTALITY RATE (%) OF
B. ARABICA SNAILS EXPOSED TO DIFFERENT
CONCENTRATIONS OF E. TERRACINA EXTRACT
72
hours
48
hours
24
hours
12
hours
6
hours
3
hours
2
hours
1
hour
Conc./
time
100 80 60.0 20.0 0.00 0.00 0.00 0.5%
100 60.0 20.0 20.0 0.00 0.00 1%
100 80.0 40.0 20.0 0.00 0.00 1.5%
100 40.0 0.00 0.00 2%
100 20.0 2.5%
100 3%
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Recently, leading scientists are doing many researches to find
alternative candidates against various diseases using herbal
flora as they considered non-toxic, almost safe and cheap as
well as the synthetic modern drugs considered not safe 100%
and the fatal side effects had increased greatly in the last years
[19]. So why, World Health Organization (WHO) had
recommended to evaluate the effectiveness of different plants
species as an alternatives drugs [20]. In this regard,
antibacterial affects of chloroform, petroleum ether,
methanol, ethanol and acetone extract gained from dried
leaves of E. prostrata, E. peplus and E. terracina on some
human pathogenic microbes had been investigated. Based on
the result, it was concluded that the extract of E. prostrata, E.
peplus and terracina possesses significant antibacterial
activities. Extraction by some solvents or from different
Euphorbia spp. showed various antibacterial activities when
compared with each other. These results, in agreement with
previous findings that indicated the most of the antimicrobial
active compounds were mainly participated in polar and non-
polar solvents such as methanol, acetone, chloroform,
petroleum ether and diethyl ether [21], [22], [23]. Pathogenic
susceptibility variation among the tested three Euphorbia
spp. are in accordance with many previous studies that
demonstrated antimicrobial efficacy were dependent on the
selected plant species, concentration or the types of extract
[24] [25], [26]. Acikgoz et al., [27] examined the
antimicrobial effect of methanol and chloroform extracts that
gained from Cladonia rangiformis Hoffm., and Cladonia
convoluta (Lamkey) and found the activity mainly depended
on the types of plant species examined. Similar report by
Cock [28] who examined the antibacterial activities of
methanolic extracts of Acacia aulacocarpa leaves and Acacia
complanta leaves and flowers against various Gram-negative
and Gram-positive bacteria and found significant inhibitory
effects. This variation may be due to that the three species of
Euphorbia had different chemicals compound. The GC-MS
analysis of the of leaves of E. peplus, E. prostrata and E.
terracina showed the presence of various chemicals
compounds that identified in the ethanol extracts of E. peplus,
E. prostrata and E. terracina, for example, the ethanol
extracts had propoxyethylamine in E. peplus, oxalic acid
diallyl ester in the E. prostrata and n-hexadecanoic acid in
the E. terracina [29]. As, it was reported that several factors
including genotype, climate, altitude, region, harvesting time
might be responsible for various chemical composition and
consequently affect with various degree the microorganisms
inhibition activity [30]. The methanol and petroleum ether
extracts gained from E. prostrata had a greater effect on
pathogenic P. aeruginosa in the form of zone of inhibition
than other solvent extracts and other investigated Euphorbia
species. Moreover, ethanol extract from E. peplus, methanol
extract from E. prostrata and ethanol extract from E.
terracina strongly inhibited the growth of K. pneumonia and
Shigella sp. than other investigated extracts. Although we
used the same solvent for each Euphorbia species various
inhibition activity were gained against the same micro-
organism, a justification that the inhibition was not affected
by the type of solvent only but mainly dependent on the plant
species itself, which makes this newly described biological
characters for species identification. The toxic effects of the
E. prostrata, E. peplus and E. terracina upon the snails may
be due to its inhibitory effects on the activities of some
important enzymes. Some authors reported a significant
changes in the activity of some important enzymes like
adenosine triphosphatase (ATPase), pyruvate kinase (PK)
pyruvate carboxy kinase (PEPCK) and lactate dehydrogenase
(LDH) in response to treatment of snails with synthetic or
plant-origin molluscicides [31], [32], [33]. Similar enzymatic
alterations may be responsible for the death of snails treated
with Euphorbia spp.
4. CONCLUSION
The present results indicate that Euphorbia spp. extracts are
a promising molluscicide and antimicrobial agents that can be
used as a natural alternative to the synthetic compounds,
however we believe that Euphorbia spp. need further
investigations to study the toxicity of various types of
extracts, the toxicity of various fractions of extract, its
stability under various environmental conditions and its
effects on other aquatic organisms.
ACKNOWLEDGMENT
Authors are thankful to the King Abdulaziz City for Science
and Technology (KACST) for the financial support under
grant number (A-I-34-280).
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