See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/267033853 Antimicrobial and antiproliferative activities of homalothecium sericeum (hedw.) schimp. extracts Article in Fresenius Environmental Bulletin · January 2011 CITATIONS READS 12 327 6 authors, including: Pinar Oztopcu Vatan Cansu Filik Iscen Eskisehir Osmangazi University Eskisehir Osmangazi University 32 PUBLICATIONS 187 CITATIONS 36 PUBLICATIONS 835 CITATIONS SEE PROFILE SEE PROFILE Selda Kabadere Semra Ilhan Eskisehir Osmangazi University Eskisehir Osmangazi University 39 PUBLICATIONS 271 CITATIONS 47 PUBLICATIONS 1,243 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: The synthesis of analogues of isophorone, their biotransformation and investigating their antimicrobial activities View project This study was supported by Eskişehir Osmangazi University Scientific Research Projects Committee (Project No: 2014-3859). View project All content following this page was uploaded by Pinar Oztopcu Vatan on 05 November 2014. The user has requested enhancement of the downloaded file. © by PSP Volume 20 – No 2a. 2011 Fresenius Environmental Bulletin ANTIMICROBIAL AND ANTIPROLIFERATIVE ACTIVITIES OF Homalothecium sericeum (Hedw.) Schimp. EXTRACTS Pinar Oztopcu-Vatan1,*, Filiz Savaroglu1, Cansu Filik-Iscen2, Selda Kabadere3, Semra Ilhan1 and Ruhi Uyar3 1 Department of Biology, Faculty of Art and Sciences, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey 2 Department of Elementary Education, Faculty of Education, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey 3 Department of Physiology, Faculty of Medicine, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey ABSTRACT of plant species [2]. Even though a few reviews concerning the biologically active chemical constituents of bryophytes The purpose of the current study was to investigate the have been published [2-4], the chemistry of bryophytes has possible antimicrobial and antiproliferative properties of been neglected for a long time for the following reasons: Homalothecium sericeum (Hedw.) Schimp. (Brachythe- it is very small morphologically, difficult to collect in large ciaceae). In our study, we first obtained 7 different extracts amounts as pure samples, difficult to identify, and consid- (methanol, chloroform, acetone, A, B, C and D) by two ered to be nutritionally useless to humans. However, the different extraction processes. The antimicrobial activity of Chinese, Europeans and North Americans have been using these extracts was then assessed by using the well diffusion bryophytes since time immemorial as a traditional medi- method against 6 bacterial and 7 fungal strains. cine to cure cuts, burns, external wounds, bacteriosis, pul- monary tuberculosis, neurasthenia, fractures, convulsions, None of the extracts influenced the growth of the fungi. scalds, uropathy, pneumonia etc. [4-7]. One of the features Although all of these extracts carried a variety of inhibitory that have helped bryophytes survive and maintain their potentials against just Pseudomonas aeruginosa, the ex- place in today’s flora is their content of biologically active tracts of acetone, A and C showed much better antibacte- compounds [1]. There is evidence confirming the antibiotic rial activities with minimum inhibitory concentrations of activity of bryophytes against prokaryotic cells and fungi 23.4, 93.8 and 375 µg/mL, respectively. Therefore, the [7, 8]. Dulger et al. [9] investigated the antimicrobial activ- possible cytotoxic activities of acetone extracts A and C of ity of methanol extracts from the aerial parts of eight dif- H. sericeum were determined by applying 3-(4,5-dimethyl- ferent mosses species with the disc diffusion method, and thiazol-2yl)- 2,5-diphenyltetrazolium bromide assay (MTT) concluded that methanol extracts possess mainly a moder- on rat glioma cell line (C6). Extract C was the most effec- ate activity against Gram positive and negative bacteria tive on C6 cell viability and, 85 and 170 µg/mL reduced and a weak antiyeast activity. Ilhan et al. [10] studied the the cell survival ratio by 39 and 86% in 24 h, respectively. antimicrobial activity of methanol and acetone extracts This study demonstrates for the first time that acetone and from Palustriella commutata and indicated that methanol C extracts from Homalothecium sericeum may be useful extract weakly affected some bacteria, but that it was as antibacterial and antiproliferative agents in the future; inactive against yeast and mould strains. In another study, hence, further purification of these extracts is anticipated. the antimicrobial activity of ethanol extracts of 15 Indian mosses was evaluated and it was found that only 7 of them were active against all the organisms tested [7]. Bodade et KEYWORDS: Bryophyta, Homalothecium sericeum, antibacterial, al. [1] investigated the antimicrobial activity of different antifungal, antiproliferative. extracts and indicated that all extracts showed varying effects against all the test fungi and bacteria. Further evi- dence was supplied by Spjut et al. [11], who tested 184 species of mosses and 23 species of liverworts for antitu- 1. INTRODUCTION mor activity and found that while the extracts of 43 spe- cies were active, those of 75 species were toxic to the test Bryophytes belong to a group of the oldest known land mice. The most activity was found in Brachytheciaceae, plants, which includes liverworts, hornworts and mosses [1]. Dicranaceae, Grimmiaceae, Hypnaceae, Mniaceae, Neck- More than 22.000 members of the mosses exist, spread out eraceae, Polytrichaceae, and Thuidiaceae. Since Homa- throughout the world, accounting for approximately 5.5% lothecium sericeum belongs to the family of Brachythe- ciaceae and for the aforementioned reasons, we chose to * Corresponding author investigate the properties of this moss. 461 © by PSP Volume 20 – No 2a. 2011 Fresenius Environmental Bulletin Cancer cells and pathogens that have gained resis- separately concentrated by rotary evaporation to dryness in tance to drug therapy are an increasing public health prob- vacuum (extract D, yield 2.44%). lem. Yet, there are a few really effective antifungal prepa- These yields from different extracts were then weighed, rations currently available for the treatment of fungal attack dissolved in dimethyl sulphoxide (DMSO) to final concen- in agriculture, an occurrence which can be economically tration of 200 mg/mL and stored at 4°C for further use. devastating. The logical progression of this knowledge would be to screen plants for any constituents that could 2.3. Test microorganisms show activity against pathogenic fungi, bacteria and/or cancer cells. To achieve this aim, the most investigated taxa The bacterial strains were recovered from long term at present are from the angiosperms group whereas very storage at –85°C in the cryobank. The bacteria were re- little data is available about other groups of plants, in par- freshed in Nutrient Broth (Merck, Germany) at 35–37°C, ticular bryophytes [5, 9, 10, 12, 13]. Therefore, we investi- and then inoculated on Nutrient Agar (Merck) plates to gated, for the first time, the possible antimicrobial poten- check the microbial purity. The moulds were refreshed in tial of Homalothecium sericeum (Hedw.) Schimp. against Malt Extract Agar (Merck) at 27°C. The strain numbers some bacterial and fungal species. In addition, only the most and sources of the acquired microorganisms are listed in effective antimicrobial extracts were tested to determine Table 1. their possible anticancer activity on a glioma cell line (C6). TABLE 1 - Bacterial and fungal strains used for antimicrobial activity test. 2. MATERIALS AND METHODS Bacterial strains Fungal strains aBacillus subtilis cAspergillus flavus NRRL B-209 ATCC 9807 2.1. Plant material bEnterococcus faecalis aAspergillus fumigatus ATCC 29212 NRRL 163 Plant material was collected from Hekimdag village on b c the Sundiken Mountains (Eskisehir, Turkey), at a height of Escherichia coli Aspergillus niger ATCC 25922 ATCC 10949 1240 m, in May 2006, and identified by Dr. F. Savaroglu, bPseudomonas aeruginosa aAspergillus parasiticus Eskisehir Osmangazi University. A voucher specimen (Sa- ATCC 27853 NRRL 465 varoglu 29) was deposited at the Herbarium of our depart- cSalmonella typhimurium dFusarium graminearum ment. ATCC 14028 (wild type) bStaphylococcus aureus dFusarium solani ATCC 25923 (wild type) d 2.2. Preparation of the extracts Geotrichum candidum Fresh gametophytic samples of H. sericeum were (wild type) a USDA, Agricultural Research Service, Peoria, IL, US; b Faculty of treated with 0.8% Tween 80 aqueous solution to remove Medicine, Department of Microbiology, Eskisehir Osmangazi Univer- epiphytic hosts normally found on the surface, extensively sity, Turkey; c Faculty of Science, Department of Biology, Anadolu washed in tap and distilled water, and dried on filter paper University, Turkey; d Faculty of Art and Science, Department of Biol- at room temperature. Extraction procedures were applied as ogy, Eskişehir Osmangazi University, Turkey ATCC: American Type Culture Collection; NRRL: Northern Regional described elsewhere [14, 15]. Extraction was carried out Research Laboratory through two different processes. At first, 10 g of the sample in powder was extracted 2.4. Determination of antimicrobial activity with 250 mL of 80% methanol, chloroform and acetone for The moss extracts were studied for their antibacterial