Antibacterial and Molluscicidal Activities of Euphorbia Spp. Grown Naturally in Aseer Region, Saudi Arabia

Vol 3, 2 (2017) 31-37

Contents lists available at Article URL KKU Journal of Basic and Applied Sciences Journal homepage: jbas.kku.edu.sa

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

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.

Keywords: B. arabica, E. peplus, E. prostrata, E. terracina.

effect on platelet aggregation, anti-inflammatory behavior, antioxidant, and 1. INTRODUCTION antimutagenicity [3], [4]. The way for the elimination of human pathogenic schistosomes in the country of Saudi Arabia Genus Euphorbia is one of the biggest groups in the flowering was mainly depend on regular chemotherapy of infected plants comprising the third largest genus and has many forms people and snail management by using synthetic agents including herbs, shrubs and trees ranging from succulent to molluscicides [5]. However, it was reported that the synthetic non-succulent plants [1]. The recorded number of species in agents molluscicides had many disadvantages [6], [7]. On the the genus of Euphorbia was about 2,160 that had been other hand, the antibacterial effect of some Euphorbia spp. had classified in the order Malpighiales and in family been studied previously, such as solvents extract obtained from Euphorbiaceae. Euphorbs plant is found in many area across Euphorbia hirta and Euphorbia heterophylla against a range the globe including the tropical, subtropical localities of Africa of clinically resistant pathogens by using broth microdilution and in the Americas and in temperate zones. Euphorbia species method and found that it could be used as natural antibacterial have their own economic importance so why the Euphorbs agents [8], [9]. The traditional uses of many herbal plants contribute to the floristic wealth of tropical and subtropical usually not validated scientifically so scientific methodology countries and for many other countries all over the world could provide valuable leads to discover new drugs and whereas the family composed of 322 genera and 8910 species expanded utilization of them in the future. It was found that [2]. Recently, many of Euphorbs plant have been used in about 119 of identified chemical compounds obtained from 91 medicine plants, for example, the stem is used for healing species are mainly used in drugs manufacture in many bronchitis, asthma, and various other lung problems and countries [10]. Combating the multidrug resistant showed anti-pyretic, analgesic, sedative, anxiolytic, inhibitory microorganism such as Staphylococcus, S. dysenteriae, S. enteritidis, S. paratyphi etc using natural product are the main Correspondence: E-mail: [email protected] P a g e | 31 Tel: o*******

KKU Journal of Basic and Applied Sciences goals of many researchers all over the world [11], [12], [13]. 24 h and the antibacterial activity was measured by measuring Aseer region, KSA, characterized by unique environmental the size of the inhibition zones [17]. condition and the landscape has great variety of Euphorbia Statistical analysis plants and many of them have not been examined yet for One-way analysis of variance (ANOVA) was applied to various aspects of biology. Consequently, this research determine statistically significant differences amongst tested focuses on the antibacterial and antischistosomal activities of extracts. A p ≤ 0.05 was considered statistically significant. three species of Euphorbia including Euphorbia prostrata Preparation of plant extracts for molluscicidal activity Aiton, Euphorbia peplus L. and Euphorbia terracina L. 0.5, 1, 1.5, 2, 2.5, and 3 grams from air-dried leaves of E. growing naturally in Aseer region, KSA as a first report. 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 2. MATERIALS AND METHODS filtered and used for the study of molluscicidal activity. Snails Collection Plant materials Adults freshwater snails Biomphalaria arabica (5-8 mm in Three species of healthy plants from species Euphorbia shell diameter) were collected from freshwater bodies in Besha namely E. prostrata, E. peplus and E. terracina were collected region, Saudi Arabia and were kept in plastic containers each from two different locations, E. terracina from El Sauda contain one liter of dechlorinated water for at least three days Mountains (2270 meters height) and E. prostrata and E. peplus for acclimatization under laboratory conditions. The water was from the garden of King Khalid University (2125 meters changed every day and the snails were fed daily on fresh height).The specimens were identified and the voucher kept in lettuce leaves. the herbarium of Biology department, Faculty of Science, Molluscicidal activity King Khalid University. Tests were performed in 24-well plates whereas the snails were Preparation of leaves extracts from the three Euphorbia spp. placed individually in the wells with about two ml of the tested for antibacterial activity concentration. To avoid the snails to be out of the wells, the 45 gms from leaves of E. prostrata, E. peplus and E.terracina wells were covered and the gap between the liquid and the were washed using distilled water and subjected to the drying cover didn't permit the snails to leave. Six groups of snails in shade for 15 days and the dried plant materials were crushed were exposed to concentrations of 0.5, 1, 1.5, 2, 2.5, and 3 % into fine particles (powder) using a mixer [14]. Extraction was of E. prostrata, E. peplus and E. terracina extracts for 1, 2, 3, accomplished by adding 20 ml from acetone, chloroform, 6, 12, 24 and 72 hours, then the snails were transferred to ethanol, methanol and petroleum ether to the half-gram (0.5 g) dechlorinated water for another 24 hours for recovery. After of air-dried powdered leaves materials. Samples were kept in the recovery period, the snails were examined with the aid of rotary shaker at 99 rpm at room temperature for 48 hours. The stereoscopic microscope to calculate the living and nonliving resultant extract was filtered using Whatman filter paper snails to measure the rate of mortality. The deaths of the snails (No.1) and placed in incubator at 49 C until the solvent was were confirmed by observation of contraction of the soft parts evaporated completely. Each extract was weighed, dissolved within the shell, absence of muscle contractions in response to in sterile dimethyl sulfoxide (DMSO) and subjected to the needle probe and change in the shell color. Control antibacterial activity test [15]. experiments were performed with dechlorinated water [18]. Test organisms used Five pathogenic organisms including Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus 3. RESULTS AND DISCUSSION aureus and Shigella sp. are used in this study. All bacterial strains were first sub-cultured in nutrient broth at 37 C for 24 Antibacterial properties of Euphorbia spp. extracts against hours. All strains were obtained from the Microbiology P. mirabilis Laboratory, Faculty of Science and Microbiology Laboratory, The in vitro antibacterial activities of acetone, chloroform, Faculty of Medicine, King Khalid University, Kingdom of ethanol, methanol and petroleum ether of dried leaves of E. Saudi Arabia. prostrata, E. peplus and E. terracina extracts against P. Screening for antibacterial activity mirabilis was evaluated using well diffusion methods. The efficacy of antibacterial activities of various solvent According to the results of the study (Table I), all solvents extract including acetone, chloroform, ethanol, methanol and extract significantly inhibited the growth of P. mirabilis. petroleum ether gained from dried leaves of E. prostrata, E. Among the 3 Euphorbia spp. tested, the acetone and ethanol peplus and E. terracina and tested against K. pneumoniae, P. leaf extracts of E. peplus showed the highest antibacterial mirabilis, P. aeruginosa, S. aureus and Shigella sp. were activity with a zone of inhibition 24.67±2.91 mm at 0.37 g/ml accomplished by using the well diffusion method [16]. 20 ml concentration. The chloroform, methanol and petroleum ether from sterilized Mueller Hinton agar (Oxoid, England) media extract of the dried leaves of E. prostrata had a slightly lower was poured into sterile Petri dish and 0.1 ml of standardized antibacterial activity than the acetone and ethanol leaf extracts inoculums of each test bacterium swapped onto agar. 6 mm of E. peplus with a zone of inhibition ranging from 24.33±0.67 diameter well was cut from the agar and each well filled with to 24.00±1.00 mm. Similarly, ethanol and petroleum ether 0.1 ml of the plant extract. Each extract was assayed in extract from E. terracina had also a slightly lower antibacterial triplicate and sterile and 0.1 ml from dimethyl sulfoxide activity with a zone of inhibition 24.33±2.33 mm and (DMSO) used as negative control and cefoxitin (30 mcg) as 24.33±0.66 mm respectively. On the other hand, the acetone positive controls. All Petri dishes were incubated at 30 C for and ethanol extract of E. prostrata and acetone, chloroform and methanol extract had a moderate antibacterial activity with P a g e | 32

KKU Journal of Basic and Applied Sciences 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 23.33±1.67 20.00±0.00* 27.00±0.58 NI (12.16-37.17) (16.16-30.50) (20.00-20.00) (24.52-29.49) 19.33±2.96** 24.33±0.67 22.33±1.45 27.00±0.58 NI (6.59-32.08) (21.46-27.20) (16.08-28.58) (24.52-29.49) 24.67±2.40 20.67±0.67* 24.33±2.33 27.00±0.58 NI (14.32-35.01) (17.70-23.53) (14.29-34.37) (24.52-29.49) 23.00±2.65 24.00±1.00 21.67±1.67* 27.00±0.58* NI (11.62-34.38) (19.60-28.30) (14.40-28.84) (24.52-29.49) 24.33±2.33 24.33±1.33 24.33±0.66 27.00±0.58 NI (14.29-34.37) (18.50-30.07) (21.46-27.20) (24.52-29.49) 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 showed modest activity with a zones of inhibition 23.00±2.89 antibacterial effect with their respective diameters of mm. Ethanol and methanol inhibition zones of 19.33±2.96 mm. No antibacterial activity extracts from E. peplus, acetone extract from E. prostrata and was observed against any bacterial for negative control. acetone and petroleum ether extracts from E. terracina showed Antibacterial properties of Euphorbia spp. extracts against a slightly lower antibacterial activity with a zone of inhibition P.aeruginosa ranging from 21.00±1.00 mm to 20.00±0.00 mm. Chloroform In Table (II), chloroform, petroleum ether, methanol, ethanol extracts from E. peplus showed lowest activity with zones of and acetone extracts of dried leaves of E. prostrata, E. peplus inhibition 17.67±2.33 mm. No antibacterial activity was and E. terracina showed varied antibacterial activity against observed against any bacterial for negative control. 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 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 E. peplus E. prostrata E. terracina P.C N.C

strains Acetone 23.00±2.89 21.66±1.66 20.00±0.00 20.33±0.67 NI (10.58-35.42) (14.40-28.84) (20.00-20.00) (17.46-23.20)

Chloroform 17.67±2.33 23.33±1.66 23.33±1.67 20.33±0.67 NI (7.63-27.71) (16.16-30.50) (16.16-30.50) (17.46-23.20)

Ethanol 21.33±1.33 20.00±0.00** 23.333±1.67 20.33±0.67 NI (15.50-27.07) (2.00-20.00) (16.16-30.50) (17.46-23.20) Methanol 21.00±1.00 26.67±0.88* 23.33±1.67 20.33±0.67 NI P. aeruginosa (16.60-25.30) (22.87-30.46) (16.16-30.50) (17.46-23.20) Petroleum ether 19.67±2.60 26.33±1.33 21.33±1.33* 20.33±0.67 NI (8.47-30.87) (20.50-32.07) (15.50-27.07) (17.46-23.20) 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 peplus with a zone of inhibition between 24.33±2.33 and S. aureus 24.00±0.67 mm. Acetone and petroleum ether extract from E. Table (III) showed that the results of solvents extracts of three prostrata and E. terracina showed a slightly lower antibacterial Euhorbia spp. against S. aureus. It inhibited by the plant activity with a zone of inhibition ranging 23.33±1.67 mm to solvent extract between 24.67±1.76 mm to 20.00±0.00 mm. 23.00±1.53 mm. Chloroform and petroleum ether extracts Ethanol extracts from E. terracina had the highest antibacterial from E. peplus showed lowest activity with zones of inhibition activities against staphylococcus with zones of inhibition of 20.00±0.00 m 24.67±1.76 mm, followed by those prepared in acetone and methanol extract form E. terracina and E.

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TABLE III: SUSCEPTIBILITY OF S. AUREUS

(20.00-20.00) (11.19-34.14) (16.16-30.50) (25.52-30.48) Ethanol 22.00±2.52 21.67±1.76* 24.67±1.76 28.00±0.58 NI

aureus

(11.17-32.83) (14.08-29.26) (17.08-32.26) (25.52-30.48)

S. Methanol 24.33±2.33** 23.00±1.53 24.00±0.67 28.00±0.58 NI (14.29-34.37) (16.43-29.57) (21.46-27.20) (25.52-30.48) Petroleum ether 20.00±0.00 23.33±1.67* 23.33±1.67 28.00±0.58 NI (20.00-20.00) (16.16-30.50) (16.16-30,50) (25.52-30.48) 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 respectively. Acetone, methanol and petroleum ether extract against Shigella sp. from E. prostrata showed a slightly lower antibacterial activity Table (IV) showed that all solvents extracts gained from dried with a zone of inhibition between 24.67±0.33 mm to leaves of E. peplus, E. peplus and E. terracina had varied 23.00±1.53 mm. Acetone, methanol and petroleum ether antibacterial activity against Shigella sp. in the range between extracts from E. terracina and chloroform, ethanol and 25.67±1.20 mm to 20.00±0.00 mm. Methanol extract from E. petroleum ether extracts from E. peplus showed lowest activity peplus and ethanol extract from E. terracina had the highest with zones of inhibition ranging from 20.67±2.96 to antibacterial activities with inhibition zone of 25.67±1.20 mm 20.00±0.00 mm. No antibacterial activity was observed and 25.00±0.00 mm against any bacterial for negative control.

TABLE IV: SUSCEPTIBILITY OF SHIGELLA SP.

(15.40-25.84) (16.42-28.92) (20.00-20.00) (26.52-31.48)

sp. Ethanol 20.67±2.96* 23.00±2.00* 25.00±0.00** 29.00±0.58 NI (7.92-33.41) (14.39-31.61) (25.00-25.00) (26.52-31.48) Methanol 25.67±1.20** 24.00±2.08** 23.33±1.67** 29.00±0.58 NI

Shigella (20.40-30.84) (15.04-32.96) (16.16-30.50) (26.52-31.48) Petroleum ether 20.00±0.00** 24.67±0.33** 21.67±1.67 29.00±0.58 NI (20.00-20.00) (23.23-26.10) (14.40-28.84) (26.52-31.48) 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 25.67±1.45 mm and 25.00±0.00 mm respectively. Acetone against K. pneumonia and methanol extracts from E. peplus, ethanol and petroleum E. peplus, E. peplus and E. terracina solvents extracts against ether extracts from E. prostrata and chloroform, methanol and K. pneumoniae had prominent antibacterial activity with petroleum ether extracts from E. terracina showed slightly less inhibition zones ranging between 25.67±1.45 mm to activity with zones of inhibition ranging from 23.33±1.67 to 21.00±3.51 mm as shown in (Table V). Ethanol extract from 23.00±1.53 mm. Chloroform extract from E. prostrata showed E. peplus and methanol extract from E. prostrata had the the lowest activity with zones of inhibition 20.00±0.00mm. highest antibacterial activities with zones of inhibition

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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 E. peplus E. prostrata E. terracina P.C N.C strains Acetone 23.33±1.67** 21.67±1.67** 21.67±1.67* 29.33±0.33 NI (16.16-30.50) (14.40-28.84) (14.40-28.84) (27.80-30.77) Chloroform 21.00±3.51 20.00±0.00* 23.33±1.67** 29.33±0.33 NI (5.89-36.11) (20.00-20.00) (16.16-30.50) (27.80-30.77) Ethanol 25.67±1.45* 23.00±1.53 21.67±1.67* 29.33±0.33 NI (19.42-31.92) (16.43-29.57) (14.40-28.84) (27.80-30.77) Methanol 23.33±3.28** 25.00±0.00* 23.33±1.67* 29.33±0.33 NI

K. K. pneumoniae (9.21-37.46) (25.00-25.00) (16.16-30.50) (27.80-30.77) Petroleum 21.67±1.67** 23.00±1.53** 23.33±1.67* 29.33±0.33 NI ether (14.40-28.84) (16.43-29.57) (16.16-30.50) (27.80-30.77) 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 concentrations 0.5 and 1% concentrations of the same extract. The present data revealed that aqueous leaves extract of E. Concerning the extract of E. prostrata, 100% mortality peplus, E. prostrata and E. terracina had a molluscicidal appeared after 1, 2, 3 hours of exposure to extract activity against B. arabica snails, the intermediate host of S. concentrations 3, 2.5 and 2%, respectively. After 6 hours mansoni in Saudi Arabia (Table VI, VII, VIII). For E. peplus exposure to concentrations 0.5, 1, 1.5 % all snails were died. extract, 100% mortality occurred after only 2 hours of snails' The extract of E. terracina achieved 100% mortality at 1, 2, 6 exposure to concentrations 2, 2.5 and 3%. For 1.5% and 48 hours of exposure to concentrations 3, 2.5, 2, 0.5% concentration, 100% mortality appeared after 3 hours of respectively. For 1 and 1.5% concentrations, 100% mortality exposure. 100% mortality delayed to 24 hours for was achieved after 24 hours of exposure.

TABLE VI: MORTALITY RATE (%) OF B. ARABICA SNAILS EXPOSED TO DIFFERENT CONCENTRATIONS OF E. PEPLUS EXTRACT Conc./ 1 2 3 6 12 24 48 72 time hour hours hours hours hours hours hours hours 0.5% 0.00 0.00 0.00 0.00 0.00 100 1% 0.00 0.00 20 20.0 20.0 100 1.5% 0.00 20.0 100 2% 40.0 100 2.5% 60.0 100 3% 100 100 TABLE VII: MORTALITY RATE (%) OF B. ARABICA SNAILS EXPOSED TO DIFFERENT CONCENTRATIONS OF E. PROSTRATA EXTRACT Conc./ 1 2 3 6 12 24 48 72 time hour hours hours hours hours hours hours hours 0.5% 0.00 0.00 0.00 100 1% 0.00 20.0 20.0 100 1.5% 0.00 20.0 40.0 100 2% 40.0 40.0 100 2.5% 60.0 100 3% 100 TABLE VIII: MORTALITY RATE (%) OF B. ARABICA SNAILS EXPOSED TO DIFFERENT CONCENTRATIONS OF E. TERRACINA EXTRACT Conc./ 1 2 3 6 12 24 48 72 time hour hours hours hours hours hours hours hours 0.5% 0.00 0.00 0.00 20.0 60.0 80 100 1% 0.00 0.00 20.0 20.0 60.0 100 1.5% 0.00 0.00 20.0 40.0 80.0 100 2% 0.00 0.00 40.0 100 2.5% 20.0 100 3% 100

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Recently, leading scientists are doing many researches to find characters for species identification. The toxic effects of the alternative candidates against various diseases using herbal E. prostrata, E. peplus and E. terracina upon the snails may flora as they considered non-toxic, almost safe and cheap as be due to its inhibitory effects on the activities of some well as the synthetic modern drugs considered not safe 100% important enzymes. Some authors reported a significant and the fatal side effects had increased greatly in the last years changes in the activity of some important enzymes like [19]. So why, World Health Organization (WHO) had adenosine triphosphatase (ATPase), pyruvate kinase (PK) recommended to evaluate the effectiveness of different plants pyruvate carboxy kinase (PEPCK) and lactate dehydrogenase species as an alternatives drugs [20]. In this regard, (LDH) in response to treatment of snails with synthetic or antibacterial affects of chloroform, petroleum ether, plant-origin molluscicides [31], [32], [33]. Similar enzymatic methanol, ethanol and acetone extract gained from dried alterations may be responsible for the death of snails treated leaves of E. prostrata, E. peplus and E. terracina on some with Euphorbia spp. human pathogenic microbes had been investigated. Based on the result, it was concluded that the extract of E. prostrata, E. 4. CONCLUSION peplus and terracina possesses significant antibacterial activities. Extraction by some solvents or from different The present results indicate that Euphorbia spp. extracts are Euphorbia spp. showed various antibacterial activities when a promising molluscicide and antimicrobial agents that can be compared with each other. These results, in agreement with used as a natural alternative to the synthetic compounds, previous findings that indicated the most of the antimicrobial however we believe that Euphorbia spp. need further active compounds were mainly participated in polar and non- investigations to study the toxicity of various types of polar solvents such as methanol, acetone, chloroform, extracts, the toxicity of various fractions of extract, its petroleum ether and diethyl ether [21], [22], [23]. Pathogenic stability under various environmental conditions and its susceptibility variation among the tested three Euphorbia effects on other aquatic organisms. 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 ACKNOWLEDGMENT [24] [25], [26]. Acikgoz et al., [27] examined the antimicrobial effect of methanol and chloroform extracts that Authors are thankful to the King Abdulaziz City for Science gained from Cladonia rangiformis Hoffm., and Cladonia and Technology (KACST) for the financial support under convoluta (Lamkey) and found the activity mainly depended grant number (A-I-34-280). on the types of plant species examined. Similar report by Cock [28] who examined the antibacterial activities of REFERENCES methanolic extracts of Acacia aulacocarpa leaves and Acacia complanta leaves and flowers against various Gram-negative [1] Govaerts, R., Fordin, D., Radcliffe-Smith, A. "World Checklist and Gram-positive bacteria and found significant inhibitory and Bibliography of Euphorbiaceae (and Pandaceae). The Royal effects. This variation may be due to that the three species of Botanic Gardens, Kew" (2000) World Checklist and Bibliography of Euphorbiaceae (and Pandaceae), pp. 1-1661. Euphorbia had different chemicals compound. 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