University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Evaluation of Methyl Red and Lactate as a Mediator and a Simple Carbon Source on Electrochemical Performance of Urmia lake Sediment Microbial Fuel Cell Evaluation of Methyl Red and Lactate as a Mediator and a Simple Carbon Source on Electrochemical Performance of Urmia lake Sediment Microbial Fuel Cell 1 10 23094 10.22108/bjm.2018.104406.1061 FA Shahnaz Mohammadi M.Sc., Dep. Biology, Faculty of Natural Sciences, University of Tabriz Tabriz, Iran Gholamreza Zarrini Associate professor, Dep. Biology, Faculty of Natural sciences, University of Tabriz,Tabriz, Iran Iraj Ahadzadeh Assistant professor, Dep. Physical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Iran Journal Article 2017 05 29 Introduction: The energy crisis is an urgent issue due to the increased consumption of fossil fuels. Therefor alternative energy sources are, of critical importance. Sediment Microbial fuel cells (SMFCs) are more important among other renewable energy sources in which chemical energy in organic compounds is converted to electrical energy due to proper bacteria (exoelectrogens) catalytic activity. Materials and methods: In this study, one liter glassy reactor was used, half of it was filled with Urmia lake sediment, where microbial consortium are present, as anodic part and upper half was filled with lake water as cathodic part. Copper wires attached to graphite electrodes 4×4 cm² (choice electrode) and via an external resistance 2/2 kΩ two sections related to each other. Electrochemical performance was evaluated by a digital voltimeter. The effectiveness of methyl red as mediator and lactate after determination of optimum concentration which is added every 15 days was evaluated. All fuel cells were studied for over 45 days of experiment. Results: The results demonstrated the mediator SMFC with power density of 7/54 mW/m² has a distinct difference with mediator-less SMFC with power density of 0.46 mW/m². The recorded power density of SMFC with lactate and mediator was 4/44 ± 1/44 mW/m². Discussion and conclusion: Sediment microbial consortia degrade available organic compounds and transfer to the anode electrode by using synthetic mediators. The results showed, in addition to external synthetic mediator, methyl red increases fuel cell electrochemical performance. While it was expected that fuel cell performs well in the presence of mediator and external carbon source, we witnessed better electrochemical performance in the absence of lactate. Introduction: The energy crisis is an urgent issue due to the increased consumption of fossil fuels. Therefor alternative energy sources are, of critical importance. Sediment Microbial fuel cells (SMFCs) are more important among other renewable energy sources in which chemical energy in organic compounds is converted to electrical energy due to proper bacteria (exoelectrogens) catalytic activity. Materials and methods: In this study, one liter glassy reactor was used, half of it was filled with Urmia lake sediment, where microbial consortium are present, as anodic part and upper half was filled with lake water as cathodic part. Copper wires attached to graphite electrodes 4×4 cm² (choice electrode) and via an external resistance 2/2 kΩ two sections related to each other. Electrochemical performance was evaluated by a digital voltimeter. The effectiveness of methyl red as mediator and lactate after determination of optimum concentration which is added every 15 days was evaluated. All fuel cells were studied for over 45 days of experiment. Results: The results demonstrated the mediator SMFC with power density of 7/54 mW/m² has a distinct difference with mediator-less SMFC with power density of 0.46 mW/m². The recorded power density of SMFC with lactate and mediator was 4/44 ± 1/44 mW/m². Discussion and conclusion: Sediment microbial consortia degrade available organic compounds and transfer to the anode electrode by using synthetic mediators. The results showed, in addition to external synthetic mediator, methyl red increases fuel cell electrochemical performance. While it was expected that fuel cell performs well in the presence of mediator and external carbon source, we witnessed better electrochemical performance in the absence of lactate.

https://bjm.ui.ac.ir/article_23094_f3c46eba9789e74245ef5a9dd33da9b1.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Investigating Synergistic Effect of Silver Nanoparticles and Nisin on Escherichia coli Genome Investigating Synergistic Effect of Silver Nanoparticles and Nisin on Escherichia coli Genome 11 23 23095 10.22108/bjm.2018.110498.1126 FA Samrand Karkon Department of Biology, Faculty of Sciences, Islamic Azad University Branch Urmia, Urmia, Iran Bahram Golestani Eimani Department of Biology, Faculty of Sciences, Islamic Azad University Branch Urmia, Urmia, Iran. 0000-0002-9134-2422 Journal Article 2018 04 28 Introduction: Considering increasing concern about the resistance of microbial infections to antibiotics and nisin peptide reducing effect (AMP) due to resistance growth in bacterial strains; extensive researches were implemented based on nanotechnology. The aim of current research was to investigate synergistic effect of silver nanoparticles conjugated with nisin on genome of Escherichia coli as a gram negative bacteria model. Materials and method: After culturing the bacteria in a Nutrient Broth medium; treatments were performed at concentrations of 50, 75, 100, 125 μg/ml of silver nanoparticles; concentrations of 25, 50, 75, 100 ,150, 200μg/ml of nisin and concentrations of 30, 50, 75 μg/ml of silver nanoparticles conjugated with nisin solution. After reading the optical density at 600 nm of control samples and treated at concentrations of 50 and 75 μg/ml of silver nanoparticles; 50 and 75 μg/ml of nisin, 50 and 75 μg/ml of silver nanoparticles conjugated with nisin, DNA was extracted and RAPD-PCR was used to investigate genomic effect. Analysis of the results of RAPD-PCR was performed by NTSYS-PC software based on Dice coefficient to calculate the similarity matrix and UPGMA. Results: The results showed that the growth inhibitory effect of silver nanoparticles conjugated with nisin was higher than of individual application of nisin and silvernanoparticles and the genomic effect of the above mentioned conjugates was higher and lower than nisin and silver nanoparticles, respectively. Therefore, the mentioned conjugated nanoparticles and nisin had no synergistic effect on the bacterial genome. Discussion and conclusion: these conjugated nanoparticles with nisin can be used as proper and strong antibacterial with the least genomic and mutation effect. Introduction: Considering increasing concern about the resistance of microbial infections to antibiotics and nisin peptide reducing effect (AMP) due to resistance growth in bacterial strains; extensive researches were implemented based on nanotechnology. The aim of current research was to investigate synergistic effect of silver nanoparticles conjugated with nisin on genome of Escherichia coli as a gram negative bacteria model. Materials and method: After culturing the bacteria in a Nutrient Broth medium; treatments were performed at concentrations of 50, 75, 100, 125 μg/ml of silver nanoparticles; concentrations of 25, 50, 75, 100 ,150, 200μg/ml of nisin and concentrations of 30, 50, 75 μg/ml of silver nanoparticles conjugated with nisin solution. After reading the optical density at 600 nm of control samples and treated at concentrations of 50 and 75 μg/ml of silver nanoparticles; 50 and 75 μg/ml of nisin, 50 and 75 μg/ml of silver nanoparticles conjugated with nisin, DNA was extracted and RAPD-PCR was used to investigate genomic effect. Analysis of the results of RAPD-PCR was performed by NTSYS-PC software based on Dice coefficient to calculate the similarity matrix and UPGMA. Results: The results showed that the growth inhibitory effect of silver nanoparticles conjugated with nisin was higher than of individual application of nisin and silvernanoparticles and the genomic effect of the above mentioned conjugates was higher and lower than nisin and silver nanoparticles, respectively. Therefore, the mentioned conjugated nanoparticles and nisin had no synergistic effect on the bacterial genome. Discussion and conclusion: these conjugated nanoparticles with nisin can be used as proper and strong antibacterial with the least genomic and mutation effect.

https://bjm.ui.ac.ir/article_23095_628178c80079fe90a7fd47fd2596fe70.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Biocomputational Investigations of Structural and Functional Properties of Cry Proteins for ‎Malaria Biocontrol Biocomputational Investigations of Structural and Functional Properties of Cry Proteins for ‎Malaria Biocontrol 25 40 23096 10.22108/bjm.2018.106941.1088 FA Mahsa Jalili-Manesh M.Sc, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran Aliakbar Haddad-Mashadrizeh Assistant professor, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran 0000-0002-4597-4103 Ali Makhdoumi Assistant professor, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran Mohammad Reza Housaindokht Professor, Department of Chemistry, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran Journal Article 2017 10 09 Introduction: Nowadays, climate changes and wide usage of chemical insecticides cause the emergence of Anopheles mosquito resistant species and environmental hazards. Therefore, biocontrol as an alternative approach provides the vivid outlook for combating these risks. Hereupon, the special attention is paid to insecticides bacteria especially Bacillus thuringiensis and protein derived from it as Cry toxins. However, scrutinizing the structural and functional characteristics of these toxins have not been much attended, so it is aimed in this study by in silico investigations. Materials and methods: For this purpose Cry4, Cry11, Cry19, Cry24 and Cry39 protein sequences were extracted from NCBIdatabase, monitoring structural and functional, physicochemical, topological features, prediction of secondary and three-dimensional modelling were carried out with programs like Interproscan, Protparam, TMHMM, Psipred and modeller9.15. Finally, their binding affinity with relevant receptors was assessed by PatchDock molecular docking program. Results: The results indicate various features of physicochemical, non-secretory and toxin accumulation. Functional monitoring has traced domains with similar functions of other Cry toxin family members. Secondary structure prediction shows the existence of helixes and sheets in similar distribution pattern with each other. In addition, the design and quality evaluation of 3D structure of all toxins were obtained in desirable level. The binding affinity assessment represents stronger binding of Cry11 and Cry24 to most of the receptors. Discussion and conclusion: In general, the results of this study, in addition to provision functional data related to Cry toxin, provide optimum practical model of capable toxins production based on multimodal protein design for biocontrol of Malaria mosquitos. Introduction: Nowadays, climate changes and wide usage of chemical insecticides cause the emergence of Anopheles mosquito resistant species and environmental hazards. Therefore, biocontrol as an alternative approach provides the vivid outlook for combating these risks. Hereupon, the special attention is paid to insecticides bacteria especially Bacillus thuringiensis and protein derived from it as Cry toxins. However, scrutinizing the structural and functional characteristics of these toxins have not been much attended, so it is aimed in this study by in silico investigations. Materials and methods: For this purpose Cry4, Cry11, Cry19, Cry24 and Cry39 protein sequences were extracted from NCBIdatabase, monitoring structural and functional, physicochemical, topological features, prediction of secondary and three-dimensional modelling were carried out with programs like Interproscan, Protparam, TMHMM, Psipred and modeller9.15. Finally, their binding affinity with relevant receptors was assessed by PatchDock molecular docking program. Results: The results indicate various features of physicochemical, non-secretory and toxin accumulation. Functional monitoring has traced domains with similar functions of other Cry toxin family members. Secondary structure prediction shows the existence of helixes and sheets in similar distribution pattern with each other. In addition, the design and quality evaluation of 3D structure of all toxins were obtained in desirable level. The binding affinity assessment represents stronger binding of Cry11 and Cry24 to most of the receptors. Discussion and conclusion: In general, the results of this study, in addition to provision functional data related to Cry toxin, provide optimum practical model of capable toxins production based on multimodal protein design for biocontrol of Malaria mosquitos.

https://bjm.ui.ac.ir/article_23096_57a74ff2d0c20c00a9d56c3a089e6669.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Using PCR-DGGE and Soil Respiration to Characterize Bacterial Changes in Heavy Metal Contaminated Soils Using PCR-DGGE and Soil Respiration to Characterize Bacterial Changes in Heavy Metal Contaminated Soils 41 56 23180 10.22108/bjm.2018.111786.1142 FA Mohammad Hossein Hemmat-Jou Department of soil science, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran 0000-0003-1317-829X Ali Akbar Safari Sanjani Department of soil science, Faculty of agriculture, Bu-Ali Sina University, Isfahan, Iran Asghar Mirzaie-Asl Department of agricultural biotechnology, Faculty of Agriculture, Bu-Ali Sina university, Hamedan, Iran Arezoo Tahmourespour Department of Basic Medical Sciences, Faculty of Medical Sciences, Isfahan (Khorasgan) branch, Islamic Azad University, Isfahan, Iran Journal Article 2018 06 25 Introduction:Soil contamination to heavy metals such as lead and zinc in and around mines causes a change in structure, complexity, diversity and activity of soil microbial communities like bacteria. Materials and methods: In the present research, PCR-DGGE approach was used to investigate the effects of Pb and Zn-contamination in Bama mine near Isfahan city on bacterial diversity, structure and complexity. Basal Respiration (BR) and Substrate Induced Respiration (SIR) was also used to assess microbial activities. Nine samples from three locations (3 for each) with different levels of heavy metal contamination were taken (from low to high), then their DNA were directly extracted. Also a 468 base pair of their 16S rRNA genes were amplified using specific primers, and their fingerprints were obtained by denaturing gradient gel electrophoresis (DGGE). BR and SIR were measured, and metabolic quotient was calculated. Finally, soil microbial activity in polluted conditions was achieved. Results: Our findings illustrate that heavy metal contamination has negative effects on bacterial diversity. By increasing the bioavailability of Pb and Zn, the complexity and diversity of bacterial communities decreased and the frequency of resistant bacteria increased. By increasing Pb and Cd contamination, SIR reduced and this shows the reduction in microbial biomass. In these conditions, SIR and metabolic quotient was more sensitive than BR, so they are better ecological indicators in polluted soils. Discussion and conclusion: Although bacterial diversity showed reduction in polluted soils, diversity is still relatively high. Bacterial ability to adapt in heavy metal contamination, bacterial resistance and their important functional roles in such conditions are valuable in soil ecosystem suggesting further researches on them. Introduction:Soil contamination to heavy metals such as lead and zinc in and around mines causes a change in structure, complexity, diversity and activity of soil microbial communities like bacteria. Materials and methods: In the present research, PCR-DGGE approach was used to investigate the effects of Pb and Zn-contamination in Bama mine near Isfahan city on bacterial diversity, structure and complexity. Basal Respiration (BR) and Substrate Induced Respiration (SIR) was also used to assess microbial activities. Nine samples from three locations (3 for each) with different levels of heavy metal contamination were taken (from low to high), then their DNA were directly extracted. Also a 468 base pair of their 16S rRNA genes were amplified using specific primers, and their fingerprints were obtained by denaturing gradient gel electrophoresis (DGGE). BR and SIR were measured, and metabolic quotient was calculated. Finally, soil microbial activity in polluted conditions was achieved. Results: Our findings illustrate that heavy metal contamination has negative effects on bacterial diversity. By increasing the bioavailability of Pb and Zn, the complexity and diversity of bacterial communities decreased and the frequency of resistant bacteria increased. By increasing Pb and Cd contamination, SIR reduced and this shows the reduction in microbial biomass. In these conditions, SIR and metabolic quotient was more sensitive than BR, so they are better ecological indicators in polluted soils. Discussion and conclusion: Although bacterial diversity showed reduction in polluted soils, diversity is still relatively high. Bacterial ability to adapt in heavy metal contamination, bacterial resistance and their important functional roles in such conditions are valuable in soil ecosystem suggesting further researches on them.

https://bjm.ui.ac.ir/article_23180_b3e2e2495f613b6036f3cb9f7a21184f.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Frequency of Mycoplasma genitalium among Patients with Bacterial Vaginosis by PCR in comparison with Culturing Method Frequency of Mycoplasma genitalium among Patients with Bacterial Vaginosis by PCR in comparison with Culturing Method 57 64 23181 10.22108/bjm.2018.108794.1100 FA Khadijeh Onsory Assistant Professor, Department of Biology, Faculty of Science, Parand Branch, Islamic Azad University, Parand, Iran Hossein Shahbani Zahiri Assistant Professor, Microbiology Department, National Genetics Research Center, Tehran, Iran Mona Abdollahi Ph.D student, Biology Department, Pardis Building, Tehran University, Tehran, Iran Zahra Haji Mehdi Nouri Ph.D student, Cellular-Molecular Department, Faculty of Science, Sirjan Branch, Islamic Azad University, Sirjan, Iran Journal Article 2018 01 02 Introduction: Mycoplasma genitalium is a vaginosis pathogen bacterium to which conventional clinical microbiology techniques cannot be applied due to difficulties in cultivation and slow growth incubation. The aim of this study was to detect the frequency of this bacteriaamong vaginosis infected women and compare it with healthy individuals using PCR and culture methods. Materials and Methods: For this purpose, we conducted a study among 150 patients with bacterial vaginosis and compared the frequency with 45 healthy women with no vaginal infections collected from Imam Zaman Hospital, Robat karim city in May up to September, 1392 (2013). To detect the cases infected with Mycoplasma genitalium, we used culture technique and Polymerase Chain Reaction (PCR) as well. Then, the sensitivity and specificity were calculated. The data was analyzed using the computer software SPSS for windows (version 19), using Cross Tab and Chi square. PResults: The results indicated that among the infected cases, 107%) 71.8) samples were positive using PCR but only 73(49.3) of samples were growing in culture. The sensitivity for culture method was reported to be 49% only while, using PCR method came to 71%. In healthy individuals, 100% of samples were negative in culture which shows specify of 100% of this technique. The specificity of PCR method was 89%. Discussion and conclusion: The findings showed that PCR is a more reliable technique to detect Mycoplasma genitalium compared to culturing method. It is also suggested that presence of this organism is strongly associated with bacterial vaginosis in female. Introduction: Mycoplasma genitalium is a vaginosis pathogen bacterium to which conventional clinical microbiology techniques cannot be applied due to difficulties in cultivation and slow growth incubation. The aim of this study was to detect the frequency of this bacteriaamong vaginosis infected women and compare it with healthy individuals using PCR and culture methods. Materials and Methods: For this purpose, we conducted a study among 150 patients with bacterial vaginosis and compared the frequency with 45 healthy women with no vaginal infections collected from Imam Zaman Hospital, Robat karim city in May up to September, 1392 (2013). To detect the cases infected with Mycoplasma genitalium, we used culture technique and Polymerase Chain Reaction (PCR) as well. Then, the sensitivity and specificity were calculated. The data was analyzed using the computer software SPSS for windows (version 19), using Cross Tab and Chi square. PResults: The results indicated that among the infected cases, 107%) 71.8) samples were positive using PCR but only 73(49.3) of samples were growing in culture. The sensitivity for culture method was reported to be 49% only while, using PCR method came to 71%. In healthy individuals, 100% of samples were negative in culture which shows specify of 100% of this technique. The specificity of PCR method was 89%. Discussion and conclusion: The findings showed that PCR is a more reliable technique to detect Mycoplasma genitalium compared to culturing method. It is also suggested that presence of this organism is strongly associated with bacterial vaginosis in female.

https://bjm.ui.ac.ir/article_23181_19f52aaf0ae4709d9b0785258b5c4b03.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Evaluating the Potential of Alkalophilic Bacteria to Enhance Concrete Durability Evaluating the Potential of Alkalophilic Bacteria to Enhance Concrete Durability 65 81 23182 10.22108/bjm.2018.110446.1124 FA Javad Hamedi Microbial Biotechnology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Tehran, Iran 0000-0002-0491-2205 Sedighe Koochakzade Microbial Biotechnology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Tehran, Iran Mohamad Saleh Labafzade Research center of Passive defence, Imam Hussein Comprehensive University Tehran, Iran Hamid Moghimi Microbial Biotechnology, College of Science, University of Tehran, Tehran, Iran Journal Article 2018 04 15 Introduction: Nowadays, concrete has been widely used as one of the best and most practical building materials in various types of structures. Concrete permeability is a factor that reduces its stability and decreases the service life of concrete structures. This study has been conducted with the aim of finding bacteria able to repair concrete surface in order to reduce its permeability. Materials and Methods: Isolated bacteria from alkaline soils of north and center of Iran were subjected to primary and secondary screening for producing urease enzyme and calcium carbonate precipitation. Selected isolates were cultured in specific broth medium for calcium carbonate precipitation. Then, the rate of their growth and calcium carbonate production were compared. Produced calcium carbonate, by the premier isolate, was verified using XRD. The ability of this isolate was evaluated in the repair of the concrete surface porosity and the reduction of its permeability by Scanning Electron Microscopy (SEM). Results: The results of primary and secondary screening showed that Bacillus sp. UTMC 2623 produces the highest amount of calcium carbonate precipitation (8.8 g/L) in three days. Its XRD analysis confirmed the presence of crystalline calcium carbonate in dried bacterial precipitate. SEM analysis showed that this bacterium creates the best coating on the concrete surface along with the calcium source. Discussion and conclusion: The present study is a report on the repairing activity of Bacillus sp. UTMC 2623 which can be used to find other native and promising microorganisms in concrete surface repair. Introduction: Nowadays, concrete has been widely used as one of the best and most practical building materials in various types of structures. Concrete permeability is a factor that reduces its stability and decreases the service life of concrete structures. This study has been conducted with the aim of finding bacteria able to repair concrete surface in order to reduce its permeability. Materials and Methods: Isolated bacteria from alkaline soils of north and center of Iran were subjected to primary and secondary screening for producing urease enzyme and calcium carbonate precipitation. Selected isolates were cultured in specific broth medium for calcium carbonate precipitation. Then, the rate of their growth and calcium carbonate production were compared. Produced calcium carbonate, by the premier isolate, was verified using XRD. The ability of this isolate was evaluated in the repair of the concrete surface porosity and the reduction of its permeability by Scanning Electron Microscopy (SEM). Results: The results of primary and secondary screening showed that Bacillus sp. UTMC 2623 produces the highest amount of calcium carbonate precipitation (8.8 g/L) in three days. Its XRD analysis confirmed the presence of crystalline calcium carbonate in dried bacterial precipitate. SEM analysis showed that this bacterium creates the best coating on the concrete surface along with the calcium source. Discussion and conclusion: The present study is a report on the repairing activity of Bacillus sp. UTMC 2623 which can be used to find other native and promising microorganisms in concrete surface repair.

https://bjm.ui.ac.ir/article_23182_2d4f2ec7d28536bd34739243efde6378.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Isolation and Identification of an Alkaliplilic Thermo-tolerant Protease producing Bacillus sp. from Dehloran Hot Spring: Production Optimization and Investigation of the Activity and Stability of the Enzyme Isolation and Identification of an Alkaliplilic Thermo-tolerant Protease producing Bacillus sp. from Dehloran Hot Spring: Production Optimization and Investigation of the Activity and Stability of the Enzyme 83 96 23216 10.22108/bjm.2018.112351.1151 FA Maryam Mehrabi Assistant professor, Department of Biology, Faculty of sciences, Razi university, Kermanshah, Iran Lale Nazari M.Sc., Department of Biology, Faculty of sciences, Razi university, Kermanshah, Iran Zhila Rostami M.Sc., Department of Biology, Faculty of sciences, Razi university, Kermanshah, Iran Journal Article 2018 08 11 Introduction:Thermophilic proteases can be used in various industries, including detergents, pharmaceuticals, food, etc. These enzymes are produced by thermophilic microorganisms, includingbacteria. Materials and methods: Sampling was carried out from Dehloran hot springs in Ilam province in the west of Iran to find new protease producing bacteria. Then, the effect of pH, temperature and finally the effect of different heating time on the production of protease enzyme were evaluated. Then, Polymerase Chain Reaction (PCR) was performed to detect the strain. Protease was purified through anion exchange using DEAE-sepharose column and its molecular weight was estimated using SDS-PAGE technique. Then, activity and stability of the enzyme were investigated in temperature and pH range. Results: Among isolated strains, bacteria Bacillus sp. DEM07 (registered in the World Gene Bank with access number KY392988) with the highest diameter of the protease clear zone, was selected to produce thermo tolerant alkaline protease. The maximum production of the alkaline protease enzyme was observed at 50 ° C, pH 7 and 48 hours after culture. The protease enzyme was purified by anionic chromatography and its molecular weight was estimated to be about 27.5 kDa after purification. The enzyme was active and stable at the temperature range of 30 to 55 ° C and the optimum temperature of the enzyme activity was observed at 50 ° C. The pH range for activity and its stability was from 4 to 11, and the optimum activity of the enzyme was observed at pH 10. Discussion and conclusion: In this study, the protease enzyme purified from Bacillus sp. DEM07 is a thermo tolerant alkalophilic protease. On the other hand, by creating the optimal conditions for achieving high production of thermo-tolerant alkalophilic protease, this enzyme has a high potential for use in various industries. Introduction:Thermophilic proteases can be used in various industries, including detergents, pharmaceuticals, food, etc. These enzymes are produced by thermophilic microorganisms, includingbacteria. Materials and methods: Sampling was carried out from Dehloran hot springs in Ilam province in the west of Iran to find new protease producing bacteria. Then, the effect of pH, temperature and finally the effect of different heating time on the production of protease enzyme were evaluated. Then, Polymerase Chain Reaction (PCR) was performed to detect the strain. Protease was purified through anion exchange using DEAE-sepharose column and its molecular weight was estimated using SDS-PAGE technique. Then, activity and stability of the enzyme were investigated in temperature and pH range. Results: Among isolated strains, bacteria Bacillus sp. DEM07 (registered in the World Gene Bank with access number KY392988) with the highest diameter of the protease clear zone, was selected to produce thermo tolerant alkaline protease. The maximum production of the alkaline protease enzyme was observed at 50 ° C, pH 7 and 48 hours after culture. The protease enzyme was purified by anionic chromatography and its molecular weight was estimated to be about 27.5 kDa after purification. The enzyme was active and stable at the temperature range of 30 to 55 ° C and the optimum temperature of the enzyme activity was observed at 50 ° C. The pH range for activity and its stability was from 4 to 11, and the optimum activity of the enzyme was observed at pH 10. Discussion and conclusion: In this study, the protease enzyme purified from Bacillus sp. DEM07 is a thermo tolerant alkalophilic protease. On the other hand, by creating the optimal conditions for achieving high production of thermo-tolerant alkalophilic protease, this enzyme has a high potential for use in various industries.

https://bjm.ui.ac.ir/article_23216_369e2f9a6543dfdf9beb2cd968b853f6.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Isolation and Identification of Fungal Endophytes of the Cowpea in Khuzestan Province Isolation and Identification of Fungal Endophytes of the Cowpea in Khuzestan Province 97 115 23253 10.22108/bjm.2018.113754.1167 FA Sahar Jonbozorgi M.Sc. Student, Plant Protection Department, Shahid Chamran University of Ahvaz, Iran Mehdi Mehrabi-Koushki Assistant professor, Department of Plant Protection, Agriculture Faculty, Biotechnology and Life Sciences Research Center, Shahid Chamran University of Ahvaz, Ahvaz, Iran Reza Farokhinejad Professor, Plant Protection Department, Shahid Chamran University of Ahvaz, Ahvaz, Iran Journal Article 2018 11 01 Introduction: Endophytes are microorganisms that colonize internal tissues of plants without causing obvious symptoms. This study was conducted to isolate and identify endophytic fungi of the cowpea in Khuzestan province. Materials and methods: During 2016, eight healthy samples of the cowpea plants were collected from the important areas under cultivation in the northern Khuzestan province. The small parts of the roots, stems, leaves and pods were deeply surface sterilized for each samples and plated on Potato-Dextrose-Agar. Sixty fungal isolates obtained in this study were purified by single spore method. Based on morphological characteristics, 21 out of 60 isolates were selected for molecular study. The isolates were grown in Potato-Dextrose-Broth and mycelial biomass was recovered by passing through filter paper. DNA extraction was performed using a phenol- and chloroform- based organic method. The parts of the nrRNA gene (ITS and 28S-D1/D2 regions) were amplified using appropriate primer pair and then sequenced. Results: The isolates were analyzedon the basis of morphological characteristics in combination with BlASTn search algorithm and ITS sequence-based phylogeny. Accordingly, the isolates were identified as follows: Alternaria destruens, Alternaria sp., Curvularia mosaddeghii, Curvularia sp., Fusarium chlamydosporum, F. nygamai, F. falciforme, F. proliferatum, Fusarium sp. Macrophomina phaseolina and Penicillium oxalicum. Discussion and conclusion: Alternaria, Fusarium and Curvularia genera were the most abundant fungal endophytes into cowpea plants, growing in warm climate of the Khuzestan Province. To the best of our knowledge, this is the first study to report endophytic growth of A. destruens, Alternaria sp., M. phaseolina, F. chlamydosporum, F. nygamai, F. falciforme, F. proliferatum and P. oxalicum within cowpea plants. Introduction: Endophytes are microorganisms that colonize internal tissues of plants without causing obvious symptoms. This study was conducted to isolate and identify endophytic fungi of the cowpea in Khuzestan province. Materials and methods: During 2016, eight healthy samples of the cowpea plants were collected from the important areas under cultivation in the northern Khuzestan province. The small parts of the roots, stems, leaves and pods were deeply surface sterilized for each samples and plated on Potato-Dextrose-Agar. Sixty fungal isolates obtained in this study were purified by single spore method. Based on morphological characteristics, 21 out of 60 isolates were selected for molecular study. The isolates were grown in Potato-Dextrose-Broth and mycelial biomass was recovered by passing through filter paper. DNA extraction was performed using a phenol- and chloroform- based organic method. The parts of the nrRNA gene (ITS and 28S-D1/D2 regions) were amplified using appropriate primer pair and then sequenced. Results: The isolates were analyzedon the basis of morphological characteristics in combination with BlASTn search algorithm and ITS sequence-based phylogeny. Accordingly, the isolates were identified as follows: Alternaria destruens, Alternaria sp., Curvularia mosaddeghii, Curvularia sp., Fusarium chlamydosporum, F. nygamai, F. falciforme, F. proliferatum, Fusarium sp. Macrophomina phaseolina and Penicillium oxalicum. Discussion and conclusion: Alternaria, Fusarium and Curvularia genera were the most abundant fungal endophytes into cowpea plants, growing in warm climate of the Khuzestan Province. To the best of our knowledge, this is the first study to report endophytic growth of A. destruens, Alternaria sp., M. phaseolina, F. chlamydosporum, F. nygamai, F. falciforme, F. proliferatum and P. oxalicum within cowpea plants.

https://bjm.ui.ac.ir/article_23253_862d576acef6af9b414f27782b0469bf.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Isolation and Identification of a Myxobacterium Producing Myxovirescins, Myxalamides and Myxochromids from Intestinal Tract of earth Worm of Ghaemshahr County Isolation and Identification of a Myxobacterium Producing Myxovirescins, Myxalamides and Myxochromids from Intestinal Tract of earth Worm of Ghaemshahr County 117 129 23281 10.22108/bjm.2018.112287.1146 FA Zahra Khosravi Babadi Ph.D student, Department of Microbiology & Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University GC, Tehran, Iran Gholam Hossein Ebrahimipour Associate professor, Department of Microbiology & Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University GC, Tehran, Iran Joachim Wink Microbial Strain Collection, Helmholtz Centre for Infection Research (HZI), Inhoffenstraße 7, D-38124 Braunschweig, Germany Journal Article 2018 08 05 Introduction: Myxobacteria are Gram-negative Deltaproteobacteria with rod-shaped vegetative cells, aerobic organotrophs that exhibit unique group behavior. Materials and Methods:The myxobacterial strain Mx18Zk was isolated from intestinal tract of earth worm of GhaemshahrCounty in Mazandaranprovince and the extraction of the fermented broth culture was prepared with organic solvent extraction method. Methanol extract tested for potential antimicrobial activity against11 various human pathogens. Finally the rest of methanol extract fractionated by HPLC and tested again against sensitive pathogens. Results: Strain Mx18Zk significantly inhibited growth of Escherichia coli, Staphylococcus aureus and Chromobacterium violaceum, therefore was used for further characterization. Analysis of morphological, biochemical, 16S rRNA gene sequence and phylogenetic tree indicated that this strain with 100 % homology belongs to the genus Corallococcus macrosporus . The methanol extract was subjected to HPLC fractionation against sensitive pathogens. The active extracts studied by LC-MS and its results compared by previously identified myxobacterial secondary metabolites deposited in Myxobase database of HZI. LC/MS analysis resulted in the identification of Myxochromids and unknown Myxovirescins, Myxalamides antibiotics in methanol extract of the strain Mx18Zk. Discussion and conclusion: On the basis of results, isolated strain was active against Escherichia coli, Staphylococcus aureus and Chromobacterium violaceum and100 % homology was belong to the genus Corallococcus macrosporus. LC/MS analysis resulted in the identification of Myxochromids and unknown Myxovirescins, Myxalamides antibiotics in methanol extract of the strain Mx18Zk. Introduction: Myxobacteria are Gram-negative Deltaproteobacteria with rod-shaped vegetative cells, aerobic organotrophs that exhibit unique group behavior. Materials and Methods:The myxobacterial strain Mx18Zk was isolated from intestinal tract of earth worm of GhaemshahrCounty in Mazandaranprovince and the extraction of the fermented broth culture was prepared with organic solvent extraction method. Methanol extract tested for potential antimicrobial activity against11 various human pathogens. Finally the rest of methanol extract fractionated by HPLC and tested again against sensitive pathogens. Results: Strain Mx18Zk significantly inhibited growth of Escherichia coli, Staphylococcus aureus and Chromobacterium violaceum, therefore was used for further characterization. Analysis of morphological, biochemical, 16S rRNA gene sequence and phylogenetic tree indicated that this strain with 100 % homology belongs to the genus Corallococcus macrosporus . The methanol extract was subjected to HPLC fractionation against sensitive pathogens. The active extracts studied by LC-MS and its results compared by previously identified myxobacterial secondary metabolites deposited in Myxobase database of HZI. LC/MS analysis resulted in the identification of Myxochromids and unknown Myxovirescins, Myxalamides antibiotics in methanol extract of the strain Mx18Zk. Discussion and conclusion: On the basis of results, isolated strain was active against Escherichia coli, Staphylococcus aureus and Chromobacterium violaceum and100 % homology was belong to the genus Corallococcus macrosporus. LC/MS analysis resulted in the identification of Myxochromids and unknown Myxovirescins, Myxalamides antibiotics in methanol extract of the strain Mx18Zk.

https://bjm.ui.ac.ir/article_23281_4dc86266ad3f894f86fd4f3cc3955416.pdf

University of Isfahan Journal of Microbial Biology 3060-7647 8 29 2019 03 21 Molecular Identification of Native Strain of Thraustochytrium 71-1 and Qualitative and Quantitative Determination of Produced Total Oil and its Squalene Molecular Identification of Native Strain of Thraustochytrium 71-1 and Qualitative and Quantitative Determination of Produced Total Oil and its Squalene 131 140 23546 10.22108/bjm.2018.110288.1119 FA Farshad Khoshbasirat Agricultural Engineering, Sciences and Modern Technologies, Graduate University of Advanced Technology, kerman, iran Shahryar Shakeri Department of Agricultural Engineering، Faculty of Sciences and Modern Technologies, Graduate University of Advanced Technology, kerman, Iran Mahmood Maleki Department of Agricultural Engineering، Faculty of Sciences and Modern Technologies, Graduate University of Advanced Technology, kerman, Iran Journal Article 2018 04 03 Introduction:Squalene is a polyunsaturated hydrocarbon with six double bonds. Squalene has many applications in biofuel, cosmetic, medical and pharmaceutical industries. The common source of Squalene is shark liver oil that its consumption is limited. Thraustochytrium a genus of unicellular marine protist belong to the thraustochytrids family which is capable of producing high Squalene. Materials and methods: In this study, the qualitative and quantitative production of Squalene, oil and biomass was investigated in the native strain of Thraustochytrium 71-1 by TLC and HPLC. Molecular characterization of this strain was analyzed by 18srDNA gene analysis. Results: Results showed that the strain 71-1 belongs to the family of Thraustochytrids and the genus of Thraustochytrium. The results showed that the strain of Thraustochytrium 71-1 produced 4.12 and 1.25 gr/L, biomass and oil, respectively. Also, Squalene was produced as 74.6 mg/L in 4 days of incubation in GYP medium. Discussion and conclusion: In this research, the production of Squalene has been reported in the strain of the thraustochytrium marine for the first time in the Middle East. It is hoped that in future, the production of this substance could be increased by optimizing the culture medium and culture conditions of this strain to produce Squalene. Introduction:Squalene is a polyunsaturated hydrocarbon with six double bonds. Squalene has many applications in biofuel, cosmetic, medical and pharmaceutical industries. The common source of Squalene is shark liver oil that its consumption is limited. Thraustochytrium a genus of unicellular marine protist belong to the thraustochytrids family which is capable of producing high Squalene. Materials and methods: In this study, the qualitative and quantitative production of Squalene, oil and biomass was investigated in the native strain of Thraustochytrium 71-1 by TLC and HPLC. Molecular characterization of this strain was analyzed by 18srDNA gene analysis. Results: Results showed that the strain 71-1 belongs to the family of Thraustochytrids and the genus of Thraustochytrium. The results showed that the strain of Thraustochytrium 71-1 produced 4.12 and 1.25 gr/L, biomass and oil, respectively. Also, Squalene was produced as 74.6 mg/L in 4 days of incubation in GYP medium. Discussion and conclusion: In this research, the production of Squalene has been reported in the strain of the thraustochytrium marine for the first time in the Middle East. It is hoped that in future, the production of this substance could be increased by optimizing the culture medium and culture conditions of this strain to produce Squalene.

https://bjm.ui.ac.ir/article_23546_5c6a82e725480900a68fc7e6d6401cc6.pdf