University of IsfahanJournal of Microbial Biology3060-76471420130120Priliminary Pages ( 1 st.Year No.4)Priliminary Pages ( 1 st.Year No.4)019493FAJournal Article20160614https://bjm.ui.ac.ir/article_19493_57c683232bf542d6bfebd1eccc3aed10.pdfUniversity of IsfahanJournal of Microbial Biology3060-76471420130120Synergistic antibacterial effects of β-Chloro-L-alanine and phosphomycin on urinary tract isolates of E. coliSynergistic antibacterial effects of β-Chloro-L-alanine and phosphomycin on urinary tract isolates of E. coli1619487FANimaHosseini JazaniAssociate Professor of Microbiology, Urmia University of Medical Sciences, IranOmidHadizadehMedical student, Student's Research Committee, Urmia University of Medical Sciences, IranHamedFarzanehMedical student, Student's Research Committee, Urmia University of Medical Sciences, IranMiladMoloudizargariVeterinary Student, Urmia University, IranJournal Article20160614 Introduction : The aim of this study was the evaluation of the synergistic effect of sub-MIC doses of Phosphomycin with β-Chloro-L-alanine against urinary tract isolates of E. coli.  Materials and method s: A total of 40 isolates were collected from urine specimens submitted to the clinical diagnostic Laboratories in Urmia, Iran. The amounts of MIC and MBC for Phosphomycin, β-Chloro-L-alanine or a mixture of 0 . 5 mM β-Chloro-L-alanine with sub-MIC doses of Phosphomycin were determined and three groups were compared.  Results : Of 40 E. coli isolates, 12. 5 % were susceptible to all investigated concentrations of phosphomycin and 2. 5 % were resistant. The mean MIC value for phosphomycin in the other E. coli isolates was determined as 25. 7± 35. 5 µg/ mL. All of the bacterial isolates were resistant to all investigated concentrations of β-Chloro-L-alanine. Application of β-Chloro-L-alanine and phosphomycin combination decreased the MIC and MBC values in 22. 5 % of the isolates.  Discussion and conclusion : This study suggests that β-Chloro-L-alanine and phosphomycin combination have in vitro synergistic effect on some isolates of urinary tract E. coli.  Introduction : The aim of this study was the evaluation of the synergistic effect of sub-MIC doses of Phosphomycin with β-Chloro-L-alanine against urinary tract isolates of E. coli.  Materials and method s: A total of 40 isolates were collected from urine specimens submitted to the clinical diagnostic Laboratories in Urmia, Iran. The amounts of MIC and MBC for Phosphomycin, β-Chloro-L-alanine or a mixture of 0 . 5 mM β-Chloro-L-alanine with sub-MIC doses of Phosphomycin were determined and three groups were compared.  Results : Of 40 E. coli isolates, 12. 5 % were susceptible to all investigated concentrations of phosphomycin and 2. 5 % were resistant. The mean MIC value for phosphomycin in the other E. coli isolates was determined as 25. 7± 35. 5 µg/ mL. All of the bacterial isolates were resistant to all investigated concentrations of β-Chloro-L-alanine. Application of β-Chloro-L-alanine and phosphomycin combination decreased the MIC and MBC values in 22. 5 % of the isolates.  Discussion and conclusion : This study suggests that β-Chloro-L-alanine and phosphomycin combination have in vitro synergistic effect on some isolates of urinary tract E. coli. https://bjm.ui.ac.ir/article_19487_9f40def9842311b0575a7e5a0e169aa0.pdfUniversity of IsfahanJournal of Microbial Biology3060-76471420130120Cloning and expression of the immunogenic moiety of Pseudomonas aeruginosa exotoxin ACloning and expression of the immunogenic moiety of Pseudomonas aeruginosa exotoxin A71419491FASaharNouri GharajelarDVSc Student of Microbiology, Urmia University, IranMalahatAhmadiAssociate Professor of Microbiology, Urmia University, IranBahmanHosseiniAssistant Professor of Biotechnology, Urmia University, IranJournal Article20160614 Introduction : Pseudomonas aeruginosa, as an opportunistic microorganism, is a major cause of nosocomial infections worldwide. Exotoxin A (ETA) is an extracellular enzyme that is produced by most clinical strains of P. aeruginosa. Although the pathogenesis of the diseases due to Pseudomonas are complex, clinical and experimental data linking ETA with the morbid and lethal consequences of Pseudomonas infection are accumulating.  Materials and method s: An immunogenic 490âbp DNA segment including translocation domain plus 1b domain of the ETA from P. aeruginosa strain PAO1 were reproduced by PCR. The PCR product was cloned in E.coli DH5α and expressed in E.coli BL21 using recombinant pET28a vector. The cloned polypeptide was found to have an electrophoretic mobility in sodium dodecyle sulfate- polyacrylamide gels (SDS -PAGE) of 18kDa.  Results : PCR and colony PCR results approved the cloning of immunogenic moiety of exotoxin A. Analysis of the location of cloned polypeptide by SDS- PAGE electrophoresis revealed that it was exported by E.coli into the bacterial periplasmic space. Discussion and conclusion : Since the whole toxin is not necessary for enhancing the immune responses and this recombinant polypeptide has antigenic qualities, so it may serve as a useful vaccine to prevent Pseudomonas infections. Introduction : Pseudomonas aeruginosa, as an opportunistic microorganism, is a major cause of nosocomial infections worldwide. Exotoxin A (ETA) is an extracellular enzyme that is produced by most clinical strains of P. aeruginosa. Although the pathogenesis of the diseases due to Pseudomonas are complex, clinical and experimental data linking ETA with the morbid and lethal consequences of Pseudomonas infection are accumulating.  Materials and method s: An immunogenic 490âbp DNA segment including translocation domain plus 1b domain of the ETA from P. aeruginosa strain PAO1 were reproduced by PCR. The PCR product was cloned in E.coli DH5α and expressed in E.coli BL21 using recombinant pET28a vector. The cloned polypeptide was found to have an electrophoretic mobility in sodium dodecyle sulfate- polyacrylamide gels (SDS -PAGE) of 18kDa.  Results : PCR and colony PCR results approved the cloning of immunogenic moiety of exotoxin A. Analysis of the location of cloned polypeptide by SDS- PAGE electrophoresis revealed that it was exported by E.coli into the bacterial periplasmic space. Discussion and conclusion : Since the whole toxin is not necessary for enhancing the immune responses and this recombinant polypeptide has antigenic qualities, so it may serve as a useful vaccine to prevent Pseudomonas infections.https://bjm.ui.ac.ir/article_19491_e43ea8724f39ae783525912c4fb1014e.pdfUniversity of IsfahanJournal of Microbial Biology3060-76471420130120Serological survey of human Toxoplasma gondii infection in northern and central regions of IranSerological survey of human Toxoplasma gondii infection in northern and central regions of Iran152019489FAKhodadadPirali-KheirabadiAssociate Professor of Parasitology, Shahrekord University, IranHosseinTahmasbyDVM, University of Shahrekord, IranKuroshManouchehri-NaeiniAssociate Professor of Medical Parasitology, Shahrekord University of Medical Sciences, IranSaeidMasoumi-GhajariJournal Article20160614<strong>Introduction</strong>: <em>Toxoplasma gondii</em> is an important zoonotic protozoan parasite that can infect man and animals. The pathogen can infect the fetus by congenital transmission during pregnancy. The aim of this study was to investigate <em>T. gondii</em> infection in people referred to health care centers in northern and central regions of Iran.
<strong>Materials and method</strong>s: Serum samples from 712 individuals in Mazandaran, Isfahan and ChaharmahalvaBakhtiari provinces, Iran, were examined for the levels of anti-<em>T. Gondii </em>IgG by ELISA. Prevalence of <em>T. gondii</em> infection in respect of gender and age was analyzed<sup>. </sup>
<strong>Results</strong>: The overall anti<strong>-</strong><em>T. gondii </em>IgG prevalence in the study population was 72.05%. In Mazandaran, Isfahan and ChaharmahalvaBakhtiari provinces,in male population respectively 87.6, 41.46 and 61.81% and in female population respectively 89.31, 47.61 and 64.44% were sero-positive with anti-<em>T. gondii </em>IgG. Sero-prevalance of anti-<em>T.gondii </em>IgG in the females was higher than in the males in the northern and central regions of Iran<sup>. </sup>
<strong>Discussion and conclusion</strong><strong>:</strong>The present study demonstrates high prevalence of <em>Toxoplasma</em> infection in northern and central regions of Iran and a higher prevalence of <em>T. gondii</em> infection was observed in females. Significant difference in infection rate between individuals living in northern and central areas in Iran was found (<em>p </em><0.05), which indicated that <em>T. gondii</em> infection is dependent on living places. Deeper investigations for the potential risk factors that threat the Iranian populations, especially female are recommended.<strong>Introduction</strong>: <em>Toxoplasma gondii</em> is an important zoonotic protozoan parasite that can infect man and animals. The pathogen can infect the fetus by congenital transmission during pregnancy. The aim of this study was to investigate <em>T. gondii</em> infection in people referred to health care centers in northern and central regions of Iran.
<strong>Materials and method</strong>s: Serum samples from 712 individuals in Mazandaran, Isfahan and ChaharmahalvaBakhtiari provinces, Iran, were examined for the levels of anti-<em>T. Gondii </em>IgG by ELISA. Prevalence of <em>T. gondii</em> infection in respect of gender and age was analyzed<sup>. </sup>
<strong>Results</strong>: The overall anti<strong>-</strong><em>T. gondii </em>IgG prevalence in the study population was 72.05%. In Mazandaran, Isfahan and ChaharmahalvaBakhtiari provinces,in male population respectively 87.6, 41.46 and 61.81% and in female population respectively 89.31, 47.61 and 64.44% were sero-positive with anti-<em>T. gondii </em>IgG. Sero-prevalance of anti-<em>T.gondii </em>IgG in the females was higher than in the males in the northern and central regions of Iran<sup>. </sup>
<strong>Discussion and conclusion</strong><strong>:</strong>The present study demonstrates high prevalence of <em>Toxoplasma</em> infection in northern and central regions of Iran and a higher prevalence of <em>T. gondii</em> infection was observed in females. Significant difference in infection rate between individuals living in northern and central areas in Iran was found (<em>p </em><0.05), which indicated that <em>T. gondii</em> infection is dependent on living places. Deeper investigations for the potential risk factors that threat the Iranian populations, especially female are recommended.https://bjm.ui.ac.ir/article_19489_a6b5e5531f66a5206a9bc02867c2d2b3.pdfUniversity of IsfahanJournal of Microbial Biology3060-76471420130120Product optimization, purification and characterization of a novel polygalacturonase produced by Macrophomina phaseolinaProduct optimization, purification and characterization of a novel polygalacturonase produced by Macrophomina phaseolina213419490FASaeedAminzadeAssistant Professor in Biochemistry, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, IranNaserFarrokhiAssistant Professor in Molecular Biology, Shahrood University, Shahrood, IranJournal Article20160614Introduction : Production of a novel polygalacturonase (PG) active at pH = 3.0 and suitable to be used in fruit juice industries from Macrophomina phaseolina was evaluated. Suitable carbon, nitrogen and phosphorous forms were determined and the condition was optimized for higher PG production.  Materials and method s: Macrophomina phaseolina was cultured in so called production medium. The secretome was separated from fungal cells and polygalacturonase was isolated via column chromatography. The biochemical activity of both secretome and isolated polygalacturonase were assayed calorimetrically. The production of polygalacturonase was optimized via changes in culture medium in terms of contents, pH and temperature and Taguchi analysis of data. Enzyme kinetics was partially performed followed by the determination of pH and temperature stability.  Results : A range of sugars except glucose and chitin demonstrated to improve the PG production. Ammonium sulfate and peptone demonstrated to be suitable nitrogen sources. Amongst phosphorous sources, dipotassium hydrogen phosphate had the greatest effect. Taguchiâs orthogonal array demonstrated that pH, temperature, ammonium sulfate and trace elements had significant effect on PG production. Furthermore, mean comparisons showed that the optimum condition achieved at pH 6.0, 35 °C, 2 g.l-1 ammonium sulfate and 2 mg.l-1 trace elements.  Discussion and conclusion : The purified PG with a relative molecular mass of 70 kDa was demonstrated its highest activity at pH = 3, and 30 °C. Amongst tested cations and chemicals, Fe2+ improved the enzyme activity by 2 fold, while the secretome responded differently.Introduction : Production of a novel polygalacturonase (PG) active at pH = 3.0 and suitable to be used in fruit juice industries from Macrophomina phaseolina was evaluated. Suitable carbon, nitrogen and phosphorous forms were determined and the condition was optimized for higher PG production.  Materials and method s: Macrophomina phaseolina was cultured in so called production medium. The secretome was separated from fungal cells and polygalacturonase was isolated via column chromatography. The biochemical activity of both secretome and isolated polygalacturonase were assayed calorimetrically. The production of polygalacturonase was optimized via changes in culture medium in terms of contents, pH and temperature and Taguchi analysis of data. Enzyme kinetics was partially performed followed by the determination of pH and temperature stability.  Results : A range of sugars except glucose and chitin demonstrated to improve the PG production. Ammonium sulfate and peptone demonstrated to be suitable nitrogen sources. Amongst phosphorous sources, dipotassium hydrogen phosphate had the greatest effect. Taguchiâs orthogonal array demonstrated that pH, temperature, ammonium sulfate and trace elements had significant effect on PG production. Furthermore, mean comparisons showed that the optimum condition achieved at pH 6.0, 35 °C, 2 g.l-1 ammonium sulfate and 2 mg.l-1 trace elements.  Discussion and conclusion : The purified PG with a relative molecular mass of 70 kDa was demonstrated its highest activity at pH = 3, and 30 °C. Amongst tested cations and chemicals, Fe2+ improved the enzyme activity by 2 fold, while the secretome responded differently.https://bjm.ui.ac.ir/article_19490_0c4ff8dcc0d265cf082a8f9a764f0f8e.pdfUniversity of IsfahanJournal of Microbial Biology3060-76471420130120Optimization of nicotinamide and riboflavin in the biodesulfurization of dibenzothiophene using response surface methodologyOptimization of nicotinamide and riboflavin in the biodesulfurization of dibenzothiophene using response surface methodology354019488FAPeymanDerikvandMSc. of Microbiology, University of Isfahan, IranZahraEtemedifarAssistant Professor of Microbiology, University of Isfahan, IranHosseinSaberMSc. of Microbial Biotechnology, University of Isfahan, IranJournal Article20160614 Dibenzothiophene (DBT) is a sulfuric compound and resistant to Hydrodesulfurization process.Rhodococcuserythropolis R1, a previously isolated bacterial strain, is capable to bioconversion of DBT to 2-hydroxybiphenyl (2-HBP) .  Materials and method s: The effect of nicotinamide (precursor of NAD) and riboflavin (precursor of FMN) on DBT biodesulfurization and growth rate by this strain was studied using Gibbs assay and turbidimeteric assay respectively. The level of cofactor precursors were optimized using response surface methodology (RSM) .  Results : Analyses showed that both nicotinamide and riboflavin were statistically significant and could enhance the biodesulfurization rate of DBT by induction of dsz operon. The optimum level of nicotinamide and riboflavin was obtained at 10.67 mM and 34.2 µM respectively .  Discussion and conclusion : In spite of increasing in BDS, the addition of these cofactor precursors led to decreased growth rate and biomass production due to limitated effect of produced 2-HBP.   Dibenzothiophene (DBT) is a sulfuric compound and resistant to Hydrodesulfurization process.Rhodococcuserythropolis R1, a previously isolated bacterial strain, is capable to bioconversion of DBT to 2-hydroxybiphenyl (2-HBP) .  Materials and method s: The effect of nicotinamide (precursor of NAD) and riboflavin (precursor of FMN) on DBT biodesulfurization and growth rate by this strain was studied using Gibbs assay and turbidimeteric assay respectively. The level of cofactor precursors were optimized using response surface methodology (RSM) .  Results : Analyses showed that both nicotinamide and riboflavin were statistically significant and could enhance the biodesulfurization rate of DBT by induction of dsz operon. The optimum level of nicotinamide and riboflavin was obtained at 10.67 mM and 34.2 µM respectively .  Discussion and conclusion : In spite of increasing in BDS, the addition of these cofactor precursors led to decreased growth rate and biomass production due to limitated effect of produced 2-HBP.  https://bjm.ui.ac.ir/article_19488_94fbfb5a1b22ee5cd13596f4bcdc38bf.pdfUniversity of IsfahanJournal of Microbial Biology3060-76471420130120The correlation of Mycolic acid production by a toluene degrading Mycobacterium in the presence of cholesterolThe correlation of Mycolic acid production by a toluene degrading Mycobacterium in the presence of cholesterol414819492FAGitiEmtiaziFull professor of Microbiology, university of Isfahan, Iran,MaryamFanaeiM.Sc. of Microbiology, University of Isfahan, Isfahan, IranAzarFarhang EsfahaniM.Sc. of Microbiology, Islamic Azad University, Falavarjan Branch, Isfahan, IranZarindokhtEmamiPh.D. student of Microbiology, Islamic Azad University, Falavarjan Branch, Isfahan, IranJournal Article20160614 Introduction : Cell wall mycolic acids (MA) from Mycobacterium tuberculosis are presented as antigens that can be used to detect antibodies as surrogate markers of active Tuberculosis (TB) disease, even in HIV coinfected patients. The use of the complex mixtures of natural MA is complicated by apparent antibody cross-reactivity with cholesterol. Mycolic acid is not only related to diagnosis of TB disease but also similar structure of mycolic acid in saprophyte strains are excellent candidate for drug delivery especially for nasal spray. The aim of this work was to investigate the effect of cholesterol on mycolic acid synthesis and potential of saprophyte mycolic acid to drug delivery.  Materials and method s: Toluene enrichment medium was used for isolation of mycolic acid producing bacteria. A fast growing, acid fast bacterium was identified by PCR reaction and the related sequence of 16S rRNA gene was deposited in the NCBI Genbank with accession number jn64433. Production of mycolic acid was investigated by high-performance liquid chromatography (HPLC) in different media.  Results : Here the effect of cholesterol on biosynthesis of mycolic acids by saprophyte Mycobacterium isolated from marine water is was reported. HPLC analyses showed the mycolic acid extracted from saprophyte. Mycobacterium had one, early, cluster of peaks, like Mycobacterium tuberculosis and Mobacterium bovis.This strain is a fast growing bacterium and cholesterol might promote longer fatty acid production. Discussion and conclusion : The similarity between mycolic acid from isolated and patogenic mycobacteria offers that mycolic acid obtained from saprophyte Mycobacterium can be useful as a drug carrier. Addition of egg yolk to media induced longer fatty acid production by isolating that is suitable for delivery of drugs into the macrophage. Introduction : Cell wall mycolic acids (MA) from Mycobacterium tuberculosis are presented as antigens that can be used to detect antibodies as surrogate markers of active Tuberculosis (TB) disease, even in HIV coinfected patients. The use of the complex mixtures of natural MA is complicated by apparent antibody cross-reactivity with cholesterol. Mycolic acid is not only related to diagnosis of TB disease but also similar structure of mycolic acid in saprophyte strains are excellent candidate for drug delivery especially for nasal spray. The aim of this work was to investigate the effect of cholesterol on mycolic acid synthesis and potential of saprophyte mycolic acid to drug delivery.  Materials and method s: Toluene enrichment medium was used for isolation of mycolic acid producing bacteria. A fast growing, acid fast bacterium was identified by PCR reaction and the related sequence of 16S rRNA gene was deposited in the NCBI Genbank with accession number jn64433. Production of mycolic acid was investigated by high-performance liquid chromatography (HPLC) in different media.  Results : Here the effect of cholesterol on biosynthesis of mycolic acids by saprophyte Mycobacterium isolated from marine water is was reported. HPLC analyses showed the mycolic acid extracted from saprophyte. Mycobacterium had one, early, cluster of peaks, like Mycobacterium tuberculosis and Mobacterium bovis.This strain is a fast growing bacterium and cholesterol might promote longer fatty acid production. Discussion and conclusion : The similarity between mycolic acid from isolated and patogenic mycobacteria offers that mycolic acid obtained from saprophyte Mycobacterium can be useful as a drug carrier. Addition of egg yolk to media induced longer fatty acid production by isolating that is suitable for delivery of drugs into the macrophage.https://bjm.ui.ac.ir/article_19492_4c81b509d2fd64ebcf5753a6b933e1e4.pdf