Eur opean Rev iew for Med ical and Pharmacol ogical Sci ences 2015; 19: 4671-4674 Antimicrobial activity of some Alnus species Ç. ALTINYAY 1, M. ERYILMAZ 2, A.N. YAZGAN 3, B. SEVER YILMAZ 3, M.L. ALTUN 3 1The Ministry of Health of Turkey, Turkish Medicines and Medical Devices Agency, Ankara, Turkey 2Department of Pharmaceutical Microbiology, Ankara University, Faculty of Pharmacy, Ankara, Turkey 3Department of Pharmacognosy, Ankara University, Faculty of Pharmacy, Ankara, Turkey Abstract. – OBJECTIVE: The increasing buted in temperate regions of the northern he - prevalence of resistant microorganisms forced misphere 1. There are six taxones of Alnus species scientists to find new antimicrobial substances growing wild in Turkey 2. The plants of the genus from different sources like medicinal plants. The aim of this study was to determine the an - Alnus contain various types of plant secondary timicrobial activities of leaf extracts of some metabolites mainly diarylheptanoids, flavonoids, Alnus sp. against some bacteria and a yeast. terpenoids, phenols, steroids and tannins 3. Alnus MATERIALS AND METHODS: Aqueous and species have been used as anti-perspirant 4, for ethanolic leaf extracts of A. glutinosa subsp. the treatment of rheumatism, uterus cancer 5, he - glutinosa, A. orientalis var. orientalis , A. orien - morrhoids 6, dental abscesses, nail inflammation 4, talis var. pubescens were screened for their an - various skin diseases such as chronic herpes, ec - timicrobial activities against Staphylococcus au - 7 6 reus ATCC 25923, S. aureus ATCC 43300 (MR - zema, prurigo and also for wound healing in SA), Bacillus subtilis ATCC 6633, Escherichia coli folk medicine. ATCC 25922, Pseudomonas aeruginosa ATCC The objective of this study was to evaluate the 27853 and Candida albicans ATCC 10231. Broth in vitro antimicrobial activities of leaf extracts of dilution method was used to determine the an - A. glutinosa subsp. glutinosa , A. orientalis var. timicrobial activities of plant extracts. orientalis , A. orientalis var. pubescens against RESULTS: Ethanolic extracts of tested species exhibited better antimicrobial activity than aque - some bacteria and a yeast . ous extracts. Ethanolic extracts of tested species possessed activity having MIC values of 0.125- 0.250 mg/ml against the tested microorganisms. Materials and Methods No antibacterial activity was observed against B. subtilis, E. coli, P. aeruginosa for all the aqueous Plant Material extracts. Except these aqueous extracts, the oth - The plant materials used in this study, their lo - ers possessed activity having MIC value of 1.000 mg/ml against the tested microorganisms. calities, collection dates and voucher specimen CONCLUSIONS: To our knowledge, this is the numbers are presented in Table I. All voucher first investigation on the evaluation of antimicro - specimens were deposited in the Herbarium of bial activities on aqueous and ethanolic leaf ex - Faculty of Pharmacy, Ankara University, Anka - tracts of these species. This study provides sig - ra, Turkey. Alnus species were identified by Prof. nificant information about antimicrobial activi - Dr. Hayri Duman. ties of leaf extracts of A. glutinosa subsp. gluti - nosa, A. orientalis var. orientalis, A. orientalis var. pubescens. It is conceivable that one of the Preparation of Extracts reason for the usage of Alnus glutinosa , in treat - 5 g of each powdered dried plant material was ment of wound healing in folk medicine, is be - extracted with water and 75% of ethanol in an ult - cause of its antimicrobial activity. rasonic bath for 1 h. The water extracts were filte - red and then lyophilized. The ethanol extracts we - Key Words: re filtered and then evaporated to dryness. Antimicrobial activity, Alnus sp., A. glutinosa, A. ori - entalis. In vitro Antibacterial and Antifungal Activities of Alnus Species Introduction Aqueous and ethanolic leaf extracts of A. glu - tinosa subsp . glutinosa , A. orientalis var . orien - The genus Alnus Miller, belongs to Betulaceae talis , A. orientalis var . pubescens were screened family, consists of about 35 species and is distri - for their potential in vitro antibacterial activities Corresponding Author: Müjde Eryılmaz, MD; e-mail: [email protected] 4671 Ç. Altınyay, M. Eryılmaz, A.N. Yazgan, B. Sever Yılmaz, M.L. Altun Table I. Scientific names, studied organs, locations, harvest dates and voucher specimen numbers of the studied plants. Voucher Studied Location Harvest specimen Scientific name organ date (AEF) No A. glutinosa subsp . glutinosa Leaves Beylice Çukurhan Village, Bolu to Sakarya May-11 25991 A. orientalis var. orientalis Leaves Köyceğiz, Sandıras Mountain, Ağla May-11 25990 A. orientalis var. pubescens Leaves Köyceğiz lakeside May-11 25989 against S. aureus ATCC 25923, S. aureus ATCC of 0.125-0.250 mg/ml against the tested microor - 43300 (MRSA), B. subtilis ATCC 6633, E. coli ganisms. No antibacterial activity was observed ATCC 25922, P. aeruginosa ATCC 27853 and against B. subtilis , E. coli , P. aeruginosa for all antifungal activities against C. albicans ATCC the aqueous extracts. The MIC values of aqueous 10231 . Prior to testing all extracts were filter-ste - extracts were 1.000 mg/ml against both S. aureus rilized through a 0.45 µm membrane filter. Broth strains and C. albicans . dilution assay was used for determination of the minimum inhibitory concentrations (MIC). The cultures were obtained in Mueller Hinton Broth Discussion (Difco, Difco Laboratories, Detroit, MI, USA ). Serial two-fold dilutions ranging from 1 .000 to A. glutinosa (L.) Gaertn., commonly known as 0.0625 mg/ml were prepared in medium. A set of “black alder” or “european alder”, is naturally tubes containing only inoculated broth were used widespread across all of Europe, from mid-Scan - as controls. After incubation for 18-24 h at dinavia to the Mediterranean countries, including 37±1° C for bacteria -48 h for fungi , the last tube northern Morocco and Algeria 10 . In Turkey, the with no microbial growth was recorded to repre - leaves of A. glutinosa have been used as a folk sent MIC value (mg/ml) 8,9 . remedy to treat wounds and cuts, and one glass of the infusion of leaves is drunk three times per day to treat rheumatism 11 . Results In the previous studies, antioxidant 12,13 , hepa - toprotective and anti-inflammatory 3 activities of The MIC results of the tested extracts are A. glutinosa have been reported . Middleton et shown in Table II . Ethanolic extracts of tested al 12 also demonstrated the antibacterial activity of species exhibited better antimicrobial activity the seed methanolic extract of A. glutinosa aga - than aqueous extracts. Ethanolic extracts of tes - inst eight bacterial species (Citrobacter freundii , ted species possessed activity having MIC values E. coli , Klebsiella aerogenes , Lactobacillus plan - Table II. MIC values (mg/ml) of Alnus species against tested microorganisms. Microorganisms Extracts S. aureus S. aureus B. subtilis E. coli P. aeruginosa C. albicans ATCC ATCC ATCC ATCC ATCC ATCC 25923 43300 6633 25922 27853 10231 A. orientalis E 0.250 0.250 0.125 0.250 0.250 0.125 var . pubescens W 1.0 1.0 -- -1.0 A. orientalis E 0.250 0.250 0.250 0.250 0.125 0.125 var. orientalis W 1.0 1.0 -- -1.0 A. glutinosa E 0.250 0.250 0.125 0.250 0.125 0.125 subsp. glutinosa W 1.0 1.0 -- -1.0 E: ethanol, W: water, ‘-‘ = represents no activity. 4672 Antimicrobial activity of some Alnus species tarum , P. aeruginosa , S. aureus , E. coli ) and the Alnus glutinosa , in treatment of wound healing in most potent activity was seen against E.coli with folk medicine, is because of its antimicrobial ac - an MIC value of 1.25x10 -1 mg/ml . In 2014, Dahi - tivity. To our knowledge, this is the first investi - ja et al 14 indicated that methanolic leaf extracts of gation on the evaluation of antimicrobial activiti - A. glutinosa possessed antimicrobial activity aga - es on aqueous and ethanolic leaf extracts of these inst B. subtilis, S. aureus, E. coli, P. aeruginosa species. and C. albicans . In 1995, Saxena et al 15 demons - trated that the bark methanolic extract of A. rub - –––––––––––––––– –-– –– ra has antibacterial effect against Gram positive Conflict of Interest and Gram negative bacteria. Diarylheptanoid The Authors declare that they have no conflict of interests. oregonin and its glycoside were identified as the two constituents responsible for this activity. In a study that the antioxidant, cytotoxic, and References antimicrobial activities of leaves, bark, and co - ne extracts of A. incana (L.) Moench ssp. inca - 1) LUDWICZUK A, S AHA A, K UZUHARA T, A SAKAWA Y. Bioac - na and A. viridis (Chaix) DC ssp. viridis were tivity guided isolation of anticancer constituents from leaves of Alnus sieboldiana (Betulaceae). evaluated, all the extracts were found to pos - Phytomedicine 2011; 18: 491-498. sess antimicrobial activity against 15 microor - 2) DAVIS , P.H. Flora of Turkey and the East Aegean ganisms. The most active extracts were found Islands, Edinburgh: Edinburg University Press as the extracts of cones of A. incana and A. vi - 2000; 11: 216. ridis with MIC values ranging from 0.117 to 3) SATI SC, S ATI N, S ATI OP. Bioactive constituents and 0.129 mg/ml 16 . medicinal importance of genus Alnus . Pharma - In a study, the ethanolic extract of the barks of cogn Rev 2011; 5: 174-183. A. pendula and the fractions obtained from it we - 4) LOI MC, P OLI F, S ACCHETTI G, S ELENU MB, B ALLERO M. re evaluated for their antibacterial activity aga - Ethnopharmacology of Ogliastra (Villagrande Stri - saili, Sardinia, Italy). Fitoterapia 2004; 75: 277- inst methicillin-resistant Staphylococcus aureus 295. (MRSA). By using the broth microdilution met - 5) HAMMOND GB, F ERNANDEZ ID, V ILLEGAS LF, V AISBERG hod, the extract found to possess antibacterial ac - AJ. A survey of traditional medicinal plants from tivity with MIC values ranging from 0.