Abdelhakim Bouyahya

Determination of Phenol Content and Antibacterial Activity of FiveMedicinal Plants Ethanolic Extracts from North-West of Morocco

The aim of this study is to screening the phytochemical content of five medicinal plants of Ouezzane province for their antibacterial activity. Ethanolic extracts were prepared using solid-liquid extraction. The total phenolic content was assessed by the Folin-Ciocalteau assay, total flavonoid content was assessed by aliminuim chloride (AlCl3) colorimetric assay. The antibacterial activity of extracts was tested against two reference strains, Escherichia coli K12 MBLA and Staphylococus aureus CECT 976 using the agar well diffusion method. The total phenol content of five ethanolic extracts ranged between 34,64 ± 1,16 and 112,48 ± 1,75 mg GAE per g extract, and the flavonoid content ranged between 9,28 ± 1,37 and 24,55 ± 0,58 mg QE/g extract. In the determination of the in vitro antimicrobial activity, flowering extracts prevented the growth of the tested strain by forming significant inhibition zones. The inhibitory activity of Ajuva Iva ethanolic extracts was especially remarkable (17,5 mm against E. coli and 21 mm against S. aureus). These species could be considered as potential sources of antibacterial compounds. Further studies are necessary for chemical characterization of the active principles and more extensive biological evaluations.

Screening of Antioxidant, Antibacterial and Antileishmanial Activities of Salvia officinalis L. Extracts from Morocco

Aims: The aim of this study was to evaluate in vitro antioxidant and antimicrobial activities of organic extracts from Salvia officinalis L. (Lamiaceae) collected in the province of Ouezzane. Study Design: Evaluation of in vitro antimicrobial and antioxidant activities of extracts and determination phenolic contents. Place and Duration of Study: After plant collection from the Province of Ouezzane, further work was carried out in Parasitology Laboratory of the National Institute of health and Laboratory of Original Research Article Et-Touys et al.; BMRJ, 16(5): 1-10, 2016; Article no.BMRJ.28307 2 Biochemistry-Immunology, Faculty of Science, Mohammed V University of Rabat, Morocco from Novembre 2015 to Mai 2016. Methodology: The antioxidant activity was evaluated using DPPH scavanging assay. The antibacterial activity was tested against three reference strains (Staphylococcus aureus, Escherichia coli and Listeria monocytogenes serovar) by the diffusion method and the minimum inhibitory concentration (MIC) by microtitration assay. The antiparasitic activity was tested against Leishmania major using MTT (3(4.5-dimethylthiazol2yl) -2.5-diphenyltetrazolium bromide) assay. The levels of polyphenols and flavonoids extracts were estimated by colorimetric assay. Results: The methanol extract has shown a significant ability to trap the radical DPPH (IC50=65.655 μg/ml) compared to n-hexane and ethanol extracts. This value is higher than that of ascorbic acid (13.198 μg/ml) and Trolox (22.484 μg/ml) used as standards. The three extracts tests revealed the inhibitory power of three bacterial strains with a significant difference in the diameters of inhibition. The largest area was registered by the hexane and methanol extracts against S. aureus (22±8 mm), while the weakest area was 11±0.22 mm expressed by the ethanol extract against E. coli and the methanol extract against L. monocytogenes. The antileishmanial activity was moderate with a value of cytotoxicity (IC50) above 1 mg / ml. The extracts showed high concentrations of polyphenols and flavonoids, while biological activities were not very high when correlated with these levels. Conclusion: These results will be completed by the determination of the active component and the extracts will be tested on other biological systems namely antifungal and antitumor activities.

In vitro Cytotoxic Effects and Antibacterial Activity of Moroccan Medicinal Plants Aristolochia longa and Lavandula multifida

Aims: The aim of this study was the evaluation of in vitro cytotoxic and antibacterial activities of organic extracts from Aristolochia longa and Lavandula multifida. Study Design: Evaluation of in vitro cytotoxic and antibacterial activities of extracts. Place and Duration of Study: Department of Biology (Faculty of Sciences), between July 2007 and July 2008. Methodology: The aerial parts were extracted by organic solvents (hexane, dichloromethane and methanol). The antibacterial activity of extracts was tested against Rhodococcus strains using the agar well diffusion method and cytotoxic activity was evaluated against three cancerous cell lines using the MTT assay. The chemical composition of extracts was determined using GC-MS. Results: The results show that the hexanic extract of A. longa (AH) and the dichloromethanic extract of A. longa (AD) present a good inhibitory effect on the three cancerous cell growth with 15 μg/ml≤IC50≤ 250 μg/ml and a total inhibitory effect on the bacterial growth with inhibition zone 30 mm at 50 mg/ml. Whereas, the extracts of L. multifida present less important inhibiting effects on the cell growth, in particular hexanic extract of L. multifida (LH) and dichloromethanic extract of Original Research Article Aneb et al.; EJMP, 16(2): 1-13, 2016; Article no.EJMP.28534 2 L. multifida (LD) with 115 μg/ml≤IC50≤ 300 μg/ml. These extracts are also active against the three strains of Rhodococcus, with more than 20 mm rings of inhibition at 50 mg/ml. The study of the chemical composition of each these species was undertaken by means of GC-MS. The phytochemical analysis of the extracts studied showed the presence of many chemical compounds which can explain these biological activities. These preliminary results suggest the presence in the extracts of compounds such as linoleic acid chloride; oleic acid and limonene-6-ol, pivalate for A. longa, and methyl linolenate; octadecane; oleic acid; 2,3,5,8-tetramethyldecane, phenol, 2methyl-5-(1-methylethyl) for L. multifida. Conclusion: This study deserves to be pursued to characterize better the asset compounds and to clarify their mechanism of action.

Determination of Phenolic Contents, Antioxidant and Antibacterial Activities of Strawberry Tree (Arbutus unedo L.) Leaf Extracts

Aims: This study was designed to evaluate the antibacterial and antioxidant activities of methanolic, ethanolic, ethyl acetate and n-hexanic extract from the leaves of Strawberry tree (Arbutus unedo L.) growing in Morocco. Study Design: Determination of total phenol and flavonoids content, in vitro antioxidant activity and antibacterial study of extracts. Place and Duration of Study: Department of Biology (Faculty of Sciences), between July 2015 and November 2015. Original Research Article Bouyahya et al.; BBJ, 14(3): 1-10, 2016; Article no.BBJ.26488 2 Methodology: The extracts were prepared using solid-liquid extraction. The total phenolic content was assessed by the Folin-Ciocalteau assay, total flavonoid content was assessed by aluminium chloride (AlCl3) colorimetric assay. The antibacterial activity of extracts was tested against five reference strains, Escherichia coli K12 MBLA, Staphylococus aureus CECT 976, Listeria monocytogenes serovar 4b CECT 4032 and Pseudomonas aeruginosa IH using the agar well diffusion method and the micro-dilution assays. The antioxidant activity was assessed by measuring the ability of the extracts to scavenge the 2,2-diphenyl-1picrylhydrazyl (DPPH). Results: The total phenol content of strawberry tree five extracts ranged between 94.51±0.08 and 141.72±0.56 mg GAE/g extract, and the flavonoid content ranged between 17.48±1.02 and 31.61±0.59 mg QE/g extract. All of the extracts presented antioxidant capacity assessed by DPPH scavenging method, but at different levels depending on the concentration and the extraction solvent are found. In addition, all extracts exhibited an antibacterial activity at different levels against strains reported as the causal agents of food borne diseases. Conclusion: Results suggest the potential use of tested A. unedo extracts as bio-preservatives in the functional food industry.

Détermination des composés phénoliques et évaluation de l’activité litholytique in vitro sur la lithiase urinaire d’extrait de Zizyphus lotus L. d’origine marocaine

RésuméZizyphus lotus L., espèce méditerranéenne, est commune au Maroc. Elle est utilisée dans la phytothérapie traditionnelle pour traiter la lithiase urinaire. Cette dernière représente au Maroc la deuxième cause d’hospitalisation en urologie après l’adénome de la prostate. Dans le but de valoriser et d’évaluer l’efficacité des fruits de Zizyphus lotus L. sur la dissolution des calculs, nous avons dosé les polyphénols totaux et les flavonoïdes puis évalué l’effet de l’extrait aqueux du fruit de Zizyphus lotus L. sur la dissolution de deux types de calculs rénaux. Les teneurs des polyphénols varient entre 34,63 et 40,49 mg EAG (équivalent acide gallique)/mg d’extrait pour l’extrait organique et 285,19 mg EAG/mg d’extrait pour l’extrait aqueux. Alors que celles des flavonoïdes varient entre 16,60 et 33,44 mg EQ (équivalent quercétine)/mg d’extrait pour l’extrait organique et 2,66 mg EQ/mg d’extrait pour l’extrait aqueux. L’extrait n-hexane présente la teneur la plus élevée, 33,44 mg EAG/mg d’extrait. Quant à l’étude in vitro, nous avons observé que l’extrait aqueux a un effet de dissolution plus important sur les calculs d’acide urique (10 %) que sur les calculs d’oxalate de calcium. Alors que l’effet du citrate et de NaCl sur la réduction de la masse des calculs est faible.AbstractZizyphus lotus L., a Mediterranean plant, is commonly found in Morocco. It is used in traditional herbal medicine to treat urolithiasis. This disease is the second cause of hospitalization in urology after prostate adenoma in Morocco. In order to develop and evaluate the effectiveness of fruit Zizyphus lotus L. on the dissolution of kidney stone, we have firstly dosed the total polyphenols and flavonoids. On the other hand, we have evolved the effect of aqueous extract of the fruit Zizyphus lotus L. on two types of kidney stones. The total phenol content of the four extracts of Zizyphus lotus L. ranged between 34.63 and 40.49 mg GAE(Gallic acid equivalent)/mg extract for organic extracts and 285.19 mg GAE/mg extract for water extracts. The flavonoid content ranged between 16.6 and 33.44 mg QE(Quercetin equivalent)/mg extract for organic extracts and 2.66 mg QE/mg extract for water extracts. Hexanic extract has the highest content of 33.44 mg GAE/mg of extract. With in vitro study, we observed that the aqueous extract has a better effect on dissolution of uric acid stones (10%) than on the calcium oxalate stones. While with citrate and NaCl, the reduction in the mass of kidney stones is small.

Origanum compactum Benth: A Review on Phytochemistry and Pharmacological Properties

Origanum compactum Benth (O. compactum) is an important aromatic and medicinal plant in Morocco. It use as a culinary condiment and largely employed in popular medicine for the treatment of ailments. Several studies recently conducted have demonstrated that O. compactum posses an antimicrobial, antifungal, antioxidant, antibacterial, anti-mutagenic, cytotoxic, anticancer, and anti-corrosion activities. Phytochemical investigations of this genus have resulted in the extraction of a number of important bioactive compounds. This emphasizes on the need of extensive study for reporting the additional information on mechanism of action of this effects.

Phytochemistry and Ethnopharmacology of Ficus carica

Ficus carica L. (Moraceae) is a plant of great importance in the traditional Arab medicine thanks to its therapeutic properties. This work was carried out in order to contribute to the ethnopharmacological knowledge of this medicinal species. We describe phytochemical compounds and ethnopharmacological properties of F. carica, species that has been used in traditional medicine for many decades. Possible trends and perspectives for future research of this plant are discussed, as well. F. carica has been found to contain several chemical constituents, mostly polyphenols and flavonoids. With its bioactive compounds, F. carica possesses a wild range of biological activities. In traditional medicine, It was reported that F. carica was applied mainly as the remedy for ulcers, indigestion and diarrhea. Some published studies have shown a broad spectrum of pharmacological activities, including antibacterial, antioxidant, antitumor, as well as anti-inflammatory activities. This paper reviews the main medicinal uses, phytochemistry and pharmacology of this plant in order to summarize its therapeutic potential and to shed light on gaps necessitating for prospected research works. Review Article Bouyahya et al.; IJBCRR, 14(1): 1-12, 2016; Article no.IJBCRR.29029 2

Analyse phytochimique et évaluation de l’activité antioxydante des échantillons du miel marocain

RésuméDans le présent travail, quatre échantillons de miel collectés des régions différentes sont analysés pour leur teneur en composés phénoliques et testés pour leur activité antioxydante. Les contenus en polyphénols totaux ont été déterminés par la méthode de Folin-Ciocalteu et les teneurs en flavonoïdes par la méthode de trichlorure d’aluminium (AlCl3). L’activité antioxydante a été évaluée par la méthode de balayage du radical 2,2-diphényl-1-picrylhydrazyl. L’estimation quantitative des flavonoïdes et des phénols totaux par la méthode colorimétrique a montré que le miel est riche en ces composés avec une différence significative entre les échantillons. Les teneurs en polyphénols s’étendent entre 124,60 ± 1,12 et 75,14 ± 0,78 mg EAG/g de miel ; alors que les pourcentages des flavonoïdes sont compris entre 43,24 ± 0,72 et 19,64 ± 0,83 mg EQ/g de miel. L’évaluation quantitative de l’effet antioxydant a montré que l’échantillon de Zoumi est le plus actif (IC50 = 48,15 μg/ml), cette valeur ne présente pas une différence significative avec les deux témoins positifs (acide ascorbique : IC50 = 27,20 μg/ml et Trolox : IC50 = 43,72 μg/ml). Nous avons trouvé une corrélation positive très élevée (R2 = 0,974) entre les teneurs en flavonoïdes et la capacité antioxydante, alors que cette corrélation est moins élevée (R2 = 0,821) entre l’activité antioxydante et les phénols totaux.AbstractIn the present work, four samples of honey collected from different regions are analyzed for phenol content and tested for their antioxidant activity. The total phenol content was determined by the Folin-Ciocalteu method and the flavonoids contents by the aluminium trichloride (AlCl3) method. The antioxidant activity was evaluated by the scanning method of radical diphenyl-1-picrylhydrazyl (DPPH). Quantitative estimation of total phenols and flavonoids content by a colorimetric method showed that honey is rich in these compounds with a significant difference between samples. The total phenol content ranges between 124.60 ± 1.12 and 75.14 ± 0.78 mg EAG/g of honey. At the same time, the percentages of flavonoids contents are between 43.24 ± 0.72 and 19.64 ± 0.83 mg EQ/g of honey. The quantitative evaluation of the antioxidant effect showed that the sample of Zoumi is most active (IC50 = 48.15 μg/ml), this value does not present a significant difference with the two positive controls (ascorbic acid: IC50 = 27.20 mg/ml and Trolox: IC50 = 43.72 μg/ml). We found a very high positive correlation (R2 = 0.974) between the flavonoids content and the antioxidant capacity, whereas this correlation was moderate (R2 = 0.821) between the antioxidant activity and the total phenol contents.

In vitro Antibacterial Activity of Organic Extracts from North-West Moroccan Medicinal Plant Myrtus communis (L.)

Aims: This study was designed to evaluate the antibacterial activity of methanol, ethanol, ethyl acetate and n-hexane extract from the leaves of Myrtus communis (L.) growing in North-West of Morocco. Study Design: In vitro evaluating of antibacterial activity and determination of total phenols and flavonoids content of extracts. Place and Duration of Study: Department of Biology (Faculty of Sciences), between July 2015 and November 2015. Methodology: The total phenol content and flavonoids content of methanol, ethanol, ethyl-acetate and n-hexane extracts were determined using Folin-Ciocalteu method and Aluminum chloride (AlCl3) colorimetric method respectively. The antibacterial activity of extracts was tested against Original Research Article Bouyahya et al.; BJI, 16(4): xxx-xxx, 2016; Article no.BJI.28500 2 five reference strains, Escherichia coli K12 MBLA, Staphylococcus aureus CECT 976, Listeria monocytogenes serovar 4b CECT 4032 and Pseudomonas aeruginosa IH using the agar well diffusion method and the micro-dilution assays. Results: The amount of the total phenolic content and total flavonoid content ranged between 86,31-137,46 gallic acid equivalents (GAE mg/g) and 14,31±1,09–31,24 quercetin equivalents (QE mg/g) of dry weight of extract, respectively. All extracts had a significant antibacterial activity against the tested bacterial strains. The biggest zones of inhibition were observed against two positive bacteria (L. monocytogenes, S. aureus). While, Gram negative bacteria (E. coli K12, P. aeruginosa) had showed to be resistant toward extracts. Conclusion: Considering these results, M. communis leaves can be used as a source of novel antimicrobial and antioxidant compounds.

Evaluation of Antimicrobial Activity of Four Organic Acids Used in Chicks Feed to Control Salmonella typhimurium: Suggestion of Amendment in the Search Standard

Today, the general public has become increasingly aware of salmonellosis problems. Organic acids are known by their antimicrobial potential and commonly used for improving the quality of poultry feed. In this context, the present work evaluated the inhibitory effect of four organic acids, namely, acetic acid, citric acid, lactic acid, and tartaric acid, at different levels of contamination by Salmonella typhimurium. The neutralization of these organic acids in vitro and in the presence of one-day-old chicks organs was also investigated during the search for Salmonella serovars in birds as described in the Moroccan standard “NM 08.0.550.” The effect of four organic acids on Salmonella typhimurium was tested in vitro and in the presence of chicks organs at different concentrations set of strain and organic acids tested. The MIC results demonstrated that tartaric acid, citric acid, and acetic acid inhibited Salmonella typhimurium at concentrations of 0.312%, 0.625%, and 0.512% for the three levels of strain: 10, 100, and 103 CFU/ml, respectively, while lactic acid and depending on the amount of the strain introduced acts differently: 0.078% for 10 CFU/ml and 0.156% for 100 and 103 CFU/ml. The concentration of 0.04M of Na2HPO4 solution has proved, in vitro, in caecums and organs of chicks (in presence of organic acids) that strain introduced, even at low concentrations, can be recovered. The use of additives has beneficial effects in Salmonella control program. However, the present results recommend the amendment of Salmonella research standard, taking into account the probable presence of organic acids in digestive content of one-day-old chicks.