Antibacterial and Molluscicidal Activities of Euphorbia

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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.

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


P a g e | 33

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.


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


NI = No inhibition; * p<0.05, ** p<0.01, represent significant difference compared with positive control

.


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


P a g e | 34

TABLE III: SUSCEPTIBILITY OF S. AUREUS



Solvent stra

ins

N.C


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



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.



Solvent stra

ins

N.C


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



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.


P a g e | 35

TABLE V: SUSCEPTIBILITY OF K. PNEUMONIAE



Solvent stra

ins

N.C


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



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


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


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%

P a g e | 36

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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Antibacterial and Molluscicidal Activities of Euphorbia