PSM Microbiology <p style="text-align: justify;">PSM Microbiology (ISSN: 2518-3834) is a peer-reviewed, multidisciplinary, open access,&nbsp; international journal that considers articles on all aspects of microbiology and allied sciences.</p> en-US (PSM Microbiology) (MANI MUGHAL) Tue, 31 Jul 2018 00:00:00 +0000 OJS 60 In vitro Studies on Biological Control of Drechslera species Causing Brown Spot Disease in Rice Plants <p style="text-align: justify;"><em>Drechslera </em>species are amongst common fungal pathogens of rice, causing leaf spot diseases. Infection with <em>Drechslera</em> species causes quantitative and qualitative damage to small grains and rice plants. In the present study, rice rhizosphere mold and yeast fungi were isolated from El-Dakahlia and El-Qaliubiya governorates.&nbsp; Results indicated that in case of mold fungi, <em>Fusarium </em>and <em>Aspergillus</em> were the most dominant isolates (520 and 410 CFU per gram, respectively) while <em>Monodictys</em> and <em>Phaeodactylium</em> represented the lowest ones (10 CFU per gram for each).&nbsp; Concerning yeast fungi, <em>Stephomoascus</em> and <em>Candida</em> gave the highest CFU count per gram (90 and 60 CFU per gram respectively). Isolation of pathogens from infected rice leaves and grains from El-Dakahlia and El-Qaliubiya governorates indicated that <em>Drechslera specifier</em> gave the highest frequency percentages (56 % and 53.33 % respectively), while <em>Drechslera rostrate</em> gave 33 % and 26.66 % for both governorates respectively.<em> Penicillium decumbens thom</em> gave the highest antagonistic activity against<em> Drechslera specifier</em> (83.9 %). Biological control of <em>Drechslera</em> species could be successfully applied by using <em>Penicillium decumbens thom </em>as a bio-agent<em>.</em></p> Naziha M Hassanein, Tahsin Shoala, Shaymaa A Gouda ##submission.copyrightStatement## Fri, 13 Jul 2018 06:32:12 +0000 Molecular Characterization of Xanthomonas oryzae pv. oryzae Isolates and its Resistance Sources in Rice Germplasm <p style="text-align: justify;">Bacterial leaf blight (BLB) of rice, the most notorious and horrendous disease of rice, caused by <em>Xanthomonas oryzae</em> pv. <em>oryzae </em>(Xoo)&nbsp; andcontinue to evolve covering more area and rice cultivars. Keeping in view the escalating virulence in pathogen population, current study was conducted to determine the genetic diversity in pathogen population and to search new sources of resistance in rice germplasm. The samples collected during the survey from different rice growing areas of Punjab were provided by the Rice Pathology group of NIAB.&nbsp; Then these samples were plated to isolate and purify the pathogen causing bacterial blight disease <em>Xanthomonas oryzae</em> pv. <em>oryzae.</em>&nbsp; The genetic diversity of the pathogens from different areas of the Punjab was determined through the repetitive DNA element with primers JEL<sup>1</sup> and JEL<sup>2</sup>. During this study, twenty seven (27) varieties/lines were screened for resistance under artificial inoculation conditions against BLB. Plants were artificially inoculated by clipping method with the <em>Xoo</em> culture. After isolation and purification of pathogen, 9 isolates were selected for genetic diversity. DNA-Fingerprinting of 9 local isolates of <em>Xoo</em> generated with IS1112 based primers Jel<sup>1</sup>and Jel<sup>2, </sup>reveal great variability among themselves. The dendrogram showed two main groups A and B. In group A isolates 1, 3 and 5 have 86% similarity while in group B isolates 9 and 8 are 100% similar and 90% similar to 7 and isolate 6 is 87% similar to 4. Where isolate 2 is 82% similar to isolates 4,6,7,8 and 9. In comparison of group A and B they are 65% similar to each other. In search of resistance source against BLB, none of the variety/line out of 27 was found immune, highly resistant, resistant or moderately resistant to bacterial leaf blight disease. Only one variety/line was moderately susceptible, five were susceptible and twenty one were highly susceptible to BLB disease. The results of this study seem to be very helpful for deploying effective management of BLB disease. The information can be utilized in controlling the bacterial leaf blight disease in Pakistan that can save the economic and production losses of rice crop that are increasing every year.</p> Ahmad Faraz Sandhu, Junaid Ahmad Khan, Safdar Ali, H M Imran Arshad, Kamran Saleem ##submission.copyrightStatement## Sun, 15 Jul 2018 00:00:00 +0000 Crude Extract of Potato Ameliorates Clinical Impacts of Infection with Helicobacter pylori: Case Study <p style="text-align: justify;">Peptic ulcer disease is a problem of global concern and the infection of <em>Helicobacter pylori</em> is the main cause of this disease. Non-pharmacological approaches to improve the outcome of this disease have been reported. The purpose of this study was to report our experience in using fresh potato juice to ameliorate clinical impacts of peptic ulcer disease in three patients who were diagnosed for <em>Helicobacter pylori</em>. They reported that after daily use of fresh potato juice for 10 days to one month, their clinical symptoms including stomach-ache, and vomiting after meal intake disappeared and accordingly their quality of life indicators including eating what they want, non-stressful episodes have been improved. Quality of life has broad meanings that include what we are deficient in, or what we cannot do due to health alterations. Peptic ulcer impacts such quality life indicators.</p> Ahed J Alkhatib ##submission.copyrightStatement## Sun, 15 Jul 2018 00:00:00 +0000 Non-classical Roles of Microbes <p style="text-align: justify;"><strong>EDITORIAL</strong></p> <p style="text-align: justify;">Classical roles of microbes include various aspects such as being involved in the etiology of diseases; here a large group of diseases are known (Costerton <em>et al</em>., 1999; Autenrieth, 2017; Wang <em>et al</em>., 2018). Other beneficial roles are also known including adherence and competition with harmful bacteria (Canny and McCormick, 2008; Hibbing <em>et al</em>., 2010). In the industrial world, microbes also have been engaged in different fields, particularly that of genetic engineering (Godwill, 2014).</p> <p style="text-align: justify;">Actually, the roles of microbes have other dimensions that may expand our horizon to understand our internal world. I would like to show our experience in the field of microbiology regarding the non-classical roles of microbes. We have found those patients with cardiac problems and who were recommended for catheterization, about 50% of them were positive for <em>Chlamydia pneumnia</em> using ELISA and PCR (Al-khatib and Al-Alawneh, 2013). Our findings and other similar findings may revolutionize the management of heart diseases.</p> <p style="text-align: justify;">Another non-classical role of microbes is crime related. We have studied 1000 prisoners in different Jordanian jails and found positive relationships between crime and latent toxoplasmosis, and according to this context, microbes impact our mental status and interfere with our perception of life (Shotar <em>et al.,</em> 2015a and b; 2016).</p> <p style="text-align: justify;">The aspects of good and bad have not become perceived from social aspects only. Our values and ethics depend indirectly on microbes. According to this context, I called these microbes as “micro-evils”, because their products are circulating in the blood and impact our nervous system (Wexler, 2007; Gandon and Vale, 2014).</p> <p style="text-align: justify;">I have recently found that <em>Candida albicans </em>expresses various important proteins including estrogen receptor, and BCL2, which implies that the pathogenicity of <em>Candida albicans </em>depends on these proteins from one side, and the functional barriers could be broken down and new concepts of interaction between microbes and host cells are likely to emerge (Alkhatib, 2017).</p> Ahed J Alkhatib ##submission.copyrightStatement## Sun, 15 Jul 2018 00:00:00 +0000 Supplemented Cell Culture Media: An Effective Approach for Enhanced Monoclonal Antibody Production <h3 style="text-align: justify;"><strong>EDITORIAL</strong></h3> <p style="text-align: justify;">Monoclonal antibodies (mAbs) are broadly used in medical research, therapeutics and diagnostics (<a href="">Jorgensen&nbsp;<em>et al.,</em>&nbsp;2007</a>). mAbs are among the best-selling class of biopharmaceutics (<a href="">Ho&nbsp;<em>et al.,</em>&nbsp;2013</a>). The in vitro hybridoma culture is an appropriate method for monoclonal antibody production using the culture supernatant (<a href="">Falkenberg&nbsp;<em>et al.,</em>&nbsp;1995</a>;&nbsp;<a href="">Sen and Roychoudhury, 2013</a>).&nbsp;The production of recombinant therapeutic biologics in cell culture is a rapidly growing industry, creating many opportunities for bioprocess engineering research and development. To meet this increasing demand for therapeutic biologics, the primary goal of increasing the yield of bioprocesses, either by increasing maximum cell densities of cultures, or improving hybridoma cell culture that requires optimizing the nutrient medium in order to support cell growth, reduce cell death, and enhance mAb production (<a href="">Kelley, 2009</a>). Supplemental feeding of amino acids has proven effective for improving cell viability (<a href="">Duval&nbsp;<em>et al.,</em>&nbsp;1991</a>) and protecting cells from apoptosis-induced death and nutrient deprivation (<a href="">Ducommun&nbsp;<em>et al.,</em>&nbsp;2001</a>).</p> <p style="text-align: justify;">The cell line and its recombinant DNA construct, culture media, and process conditions are known to have a significant effect on the expression and stability of therapeutic products. Perhaps, the most important and crucial step in cell culture, however, is the selection of appropriate growth medium for&nbsp;<em>in vitro</em>cultivation (<a href="">Zarei&nbsp;<em>et al.,</em>&nbsp;2014</a>). The cell culture medium provides an artificial environment conducive to survival and proliferation of cultured cells and maintains the desired pH and osmolality.&nbsp;The choice of cell culture media has been known to significantly affect the physiochemical characteristics of mAbs. The selection of the media depends on the type of cells to be cultured and also the purpose of the culture and resources.&nbsp;The complexity of composition of cell-culture media offers many challenges towards optimization of the individual media components. Synthetic or chemically defined media are prepared by adding defined concentration of nutrients (both organic and inorganic) such as vitamins, salts, traces elements, carbohydrates, and cofactors.</p> <p style="text-align: justify;">CHO are supplied largely in the form of glucose as a carbon source for the hybridoma cells. Besides glucose is the energy source and supplementation with glucose supports viable cells in extended stationary phase. In some cases, glucose is substituted with other sugars to improve cell growth, viability and protein production (<a href="">Wilkens<em>&nbsp;et al.,</em>&nbsp;2011</a>). Many previous studies focused on glucose and glutamine feeding to progress a strategy for procuring improved cell performance (<a href="">Glacken&nbsp;<em>et al.,</em>&nbsp;1986</a>). In numerous researches, glutamine supplementation did not stimulate cell proliferation (<a href="">Geaugey&nbsp;<em>et al.,</em>&nbsp;1989</a>) or show a reasonable growth supportive effect (<a href="">Franek, 1995</a>).</p> <p style="text-align: justify;">In our study, four test media containing RPMI-1640, 10% FBS, and penicillin/streptomycin, supplemented with glucose (control), fructose, galactose and maltose at concentration of 15 mg/mL each as carbon sources were used for cell culture. Cells were cultured in a humidified atmosphere of 5% CO<sub>2</sub>&nbsp;at 37°C to optimize media components. Spleen cells were isolated from the immunized mice and mixed with Sp2/0 murine myeloma cells at ratio of 1:10 in presence of 1mL 50% PEG. Cell fusion and hybridoma culture procedures were carried out essentially following previous reports by limiting dilution (<a href="">Iqbal&nbsp;<em>et al.</em><em>,&nbsp;</em>2018</a>;&nbsp;<a href="">Ling&nbsp;<em>et al.,</em>&nbsp;2014</a>;&nbsp;<a href="">Ling&nbsp;<em>et al.,&nbsp;</em>2018</a>;&nbsp;<a href="">Ling&nbsp;<em>et al.</em><em>,</em>&nbsp;2015</a>). The most effective culture medium for hybridoma clones was supplemented with maltose that achieved best titer as determined by icELISA. Viable hybridoma cell densities were increased when supplemented with maltose as compared to the control. The monoclonal antibody produced by using supplemented culture media was specific. Another study evaluated the use of maltose supplementation in the production of a recombinant monoclonal antibody, demonstrating improvements in recombinant monoclonal antibody titer (<a href="">Leong&nbsp;<em>et al.,&nbsp;</em>2018</a>). Therefore, the use of supplemented cell culture media is an effective approach for enhanced production of monospecific-antibody, better hybridoma growth and viability.</p> Muhammad Naeem Iqbal, Asfa Ashraf, Shihua Wang ##submission.copyrightStatement## Tue, 31 Jul 2018 00:00:00 +0000