Determining of fungal diversity with conventional and molecular methods at extreme acidic environments such as acidic mine drainage

New Biotechnology · Volume 29S · September 2012

Possible potential increases day by day when identification of new compounds and pathways, molecular and biochemical characterization of cellular components are carried out. In this study, microbial communities of Nas¸a thermal spring in Kütahya-Simav have been studied a combination of classical microbiology (cultivation techniques) such as improvement of pure culture and molecular biology approaches including fluorescent in situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE). Afterwards microbial identification of isolated microorganisms molecular methods have been carried out. Keywords: Thermophilic microorganisms; Microbial diversity; Thermal spring http://dx.doi.org/10.1016/j.nbt.2012.08.635 Poster 5.0.196 Determining of fungal diversity with conventional and molecular methods at extreme acidic environments such as acidic mine drainage Pınar Aytar1,∗ , Serap Gedikli1 , Erc¸in Kocabıyık1 , Bükay Yenice ¸ abuk2 , Semra I˙ lhan2 Gürsu2 , Mehmet Burcin Mutlu3 , Ahmet C 1

Graduate School of Natural and Applied Sciences, Eskisehir Osmangazi University, 26480, Eskisehir, Turkey 2 Department of Biology, Faculty of Arts and Science, Eskisehir Osmangazi University, 26480, Eskisehir, Turkey 3 Department of Biology, Faculty of Science, Anadolu University, Eskisehir, Turkey Acid mine drainage (AMD) is produced when sulfide-bearing material is exposed to oxygen and water and has lower acidic pH values, high specific conductivity. Several active or abandoned mines are located in Turkey, therefore there are many AMDs and very little is known about microbiological diversities of acidic environments in Turkey. The aim of the research is determining the acidophilic/acidotolerant fungal diversity living in acidic mine waters which are anthropogenic origin places such as AMDs of C ¸ an (C ¸ anakkale) (pH 2.85) and Balya (Balıkesir) (pH 2.75). Each AMD samples of 20 ml were filtered through a 0.45 ␮m membrane. Filters were placed onto four different media which were malt extract agar having different pHs, dichloran rose bengal chloramphenicol agar, dichloran 18% glycerol agar and ferrous sulphate agar. The plates were incubated for 30 days at 25 ◦ C. For each medium, average CFU were calculated at incubation days of 3, 5, 7, 14 and 30th. Pure cultures isolated from all media were identified using traditional and molecular methods. Genomic DNAs of selected filamentous microfungi were extracted using CTAB protocol. The ITS1-5,8S rDNA–ITS2 region was amplified using a combination of primers ITS1 and ITS4. Sequence similarities were obtained using BLAST tool from National Center for Biotechnology Information. Phylogenetic trees were constructed with the Neighbor-Joining method using Mega 4.0 software. As a result of this study, presence of species belonging to genera such as Aspergillus, Penicillium, Paecilomyces and Trichoderma was determined. Keywords: Acidophile; Fungus identification; Acidic mine drainage S226

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http://dx.doi.org/10.1016/j.nbt.2012.08.636 Poster 5.0.197 Lead and nickel biosorption with a dried fungal biomass isolated from lead mine drainage using response surface methodology Pınar Aytar1,3,∗ , Serap Gedikli1,3 , Yeliz Buruk2,3 , Ahmet Cabuk1,3 , Nimetullah Burnak2,3 1

Graduate School of Natural and Applied Sciences, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey 2 Department of Biology, Faculty of Arts and Science, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey 3 Department of Industrial Engineering, The Faculty of Engineering and Architecture, Eskisehir Osmangazi University, 26480 Eskisehir, Turkey Biosorption of metal ions is possibly attractive technology for treatment of acid mine drainage for separation or recovery of these ions. The present study was performed to optimize the various metal biosorption parameters using a fungal biomass isolated from lead and zinc mine drainage in Balya (Balıkesir province/Turkey). Lead and nickel concentrations of this acidic mine drainage were 1.7 and 0.22 mg/l, respectively The Box–Behnken design was used to statistically analyze the experiments and to evaluate the effects of process parameters for optimization of Pb (II) and Ni removal (%). By using Box-Behnken Design contained 29 experiments for each metal ion, the effects of independent variables: pH, initial metal concentration, contact time and biosorbent dosage were investigated. The best-fitting mathematical models of the responses were selected based on the comparisons of several statistical parameters including coefficient of determination (R2 ), the adjusted coefficient of determination (adj-R2 ) and the F-value provided by analysis of variance (ANOVA). The model summary statistics showed that the regression coefficient is best for the quadratic model. The experimental values were found to be reasonably close to the predicted ones, which confirmed the validity and adequacy of the predicted models. Furthermore, zeta potential measurements and Fourier transform infrared spectroscopy were carried out to understand the adsorption mechanism. The results of the study showed that the fungal biomass from acidic mine drainage as a biosorbent for recovery/separation of Pb(II) and Ni(II) from acid mine drainages had a significant potential. Keywords: Biosorption; Response surface methodology; Optimization; Acid mine drainage http://dx.doi.org/10.1016/j.nbt.2012.08.637