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Isolation and Molecular Characterization of Persistent Organic Pollutants Metabolizing Bacteria

Thesis Info

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Author

Muccee, Fatima

Program

PhD

Institute

The Islamia University of Bahawalpur

City

Bahawalpur

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/11741/1/Fatima%20muccee%20biotechnology%202019%20iub%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726537118

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Present study was designed to isolate and characterize the persistent organic pollutants (POPs) metabolizing bacteria from tannery effluents and petrol contaminated soil samples collected from gasoline stations of Bahawalpur, Pakistan. For the isolation of individual bacteria capable of metabolizing different POPs methods of selective enrichment culture and serial dilution were employed. Followed by isolation, growth behavior, corresponding POPs removal efficiencies, biochemical characteristics and antibiotics resistance profiles were documented. For molecular analysis, 16S rRNA gene was amplified and sequenced. Retrieved sequences were analyzed through BLAST analysis and to reflect relationship phylogenetic trees were constructed using software MEGA version 7. To detect the possible metabolic pathways responsible for POPs degradation, the isolates with high removal efficiencies were selected. Extracellular and intracellular metabolites of the selected bacteria were extracted and subjected to GC-MS and LC-MS analysis. Effect of selected carbon source on proteins production of bacteria was elucidated through LC-MS analysis of the intracellular proteins extracted from various bacterial isolates. The 16S rRNA metagenome sequencing was performed in order to confirm the POPs metabolizing potential of isolated bacteria among the bacterial community of tannery effluent and petrol contaminated soil. Shotgun metagenome sequencing helped to determine the possible genes and pathways associated with POPs degradation. Total forty-one bacteria were isolated during present study of which twenty-eight were toluene metabolizing, four were naphthalene metabolizing and nine were petrol metabolizing bacteria. The top BLAST similarity analysis revealed similarity of isolates with Brevibacillus agri strain NBRC 15538 (n = 27), Bacillus paralicheniformis strain KJ-16 (n = 4), Burkholderia lata strain 383 (n = 6), Burkholderia pyrrocinia strain LMG 14191 (n = 1) and Brevibacillus formosus strain NBRC 15716 (n = 3). All isolates were found sensitive to teicoplanin and linezolid and resistant to oxacillin and chloramphenicol. However, two bacteria, IUBT9 and IUBT26, exhibited sensitivity to oxacillin. Present study bacteria were examined to have considerable toluene removal potential (28-93%), naphthalene removal potential (70-83%) and petrol removal potential (41-69.5%). Biochemical characterization revealed presence of many valuable enzymes like arginine dehydrolase, naphthylamidase, glucosaminidase and phosphatase and ability of isolates to ferment variety of sugars like esculin, mannitol, sorbitol and inulin. GC-MS and LC-MS based analysis of bacterial metabolites confirmed degradation potential of the isolated bacteria to metabolize toluene, naphthalene and petrol through not only the earlier reported metabolic pathways but also some novel metabolic pathways which have not been reported so far. LC-MS analysis of bacterial proteins revealed the presence of enzymes involved in the degradation of toluene, synthesis of secondary metabolites, central dogma of gene expression, chemotaxis, spore development, transport of ions and central carbon metabolism further confirming the utilization of the provided carbon sources (toluene, naphthalene, petrol) by present study bacteria.
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اظہارِ تشکر

اظہارِ تشکر
اقرار مصطفیٰ
اللہ تبارک و تعالیٰ کے نہایت فضل و کرم اور نبیِ مکرمﷺ کے وسیلہ ٔ جمیلہ کے سبب نعتیہ مجموعہ’’ حسنِ کُن‘‘(مسدسِ مصطفیٰؐ)مسدس کی صورت میںمکمل ہوا۔ عہدِ رواں انتشار و خلفشار اور طرح طرح کے زوالی فتنوں کا عہد ہے۔ مسائل کی فراوانی اور وسائل کی کم یابی کے باعث ضروریات ِ حیات کو پورا کرنا وبالِ جان بن چکا ہے۔ ایسے میں خواہشات کی تکمیل کے لیے پر تولنا ، کم فہمی اور دیوانے کے خواب کے مترادف ہے۔مگر کچھ خواہشات ،ضروریات ِ حیات سے بھی بڑھ کے ہوتی ہیں کیوں کہ ان کا تعلق ایمان ومحبت سے جڑا ہوتا ہے اور اہلِ ایمان و محبت کا اعتقاد و یقین سے بھرپور او رمسلم نظریہ یہ ہے کہ تعلقِ خاطر خالصتاً نبی ِ آخرالزماں ﷺ سے ہو تو خواہشوں کی تکمیل کے خواب کو تعبیر میں ڈھلنے کے اسباب خود بہ خود پیدا ہوجاتے ہیں یا اسباب پیدا کر دیے جاتے ہیں۔ موجودہ نامساعد حالات کے پیشِ نظر ، مجھ ایسے کم مایا کا مجموعہ منصہ شہود پر آنا درحقیقت احمد رضاؒ خاں بریلوی کے اس شعر کے مصداق ہے:
کون دیتا ہے دینے کو منہ چاہیے
دینے والا ہے سچا ہمارا نبیؐ
عزیزانِ من! حُسنِ کُن‘‘مسدس کی ہئیت میں لکھا گیا ہے۔ اسمِ محمدﷺ اور اسمِ احمدﷺ کے اعداد کا مجموعہ 145بنتا ہے، اسی نسبت سے مسدس کے بندوں کی تعداد 145رکھی گئی ہے اور اسی وجہ کے پیشِ نظر اس میں خصوصی طور پہ یہ التزام ملحوظ رکھا گیا ہے کہ یہ موضوع ، موضوعِ نعت ہے اور یہ سراپا نعت ہی رہے۔
الحمد للہ!’’ حُسنِ کُن‘‘ (مسدسِ مصطفیٰؐ) حسنِ نعت سے مزین ہے۔ پھر بھی یہ بات اپنی جگہ صد فی صد درست ہے کہ:
حق تو یہ ہے کہ حق ادا نہ ہوا
خواہش اور دعا...

Relationship Between Level of Educational Attainment and Employee Performance: Mediating Role of Individual Religious Affiliation

Purpose: This study empirically investigates the relation between education level and employees’ performance working in public sector universities from the viewpoint of Islamic religiosity when religious affiliations play mediating role. It has been experienced that more educated people are found to be more involved in performing tasks related to the religious affiliations. Due to the fact that people with higher education found less time to complete their religious obligations, they are more involved in alternative arrangements like charity and donations to fulfill the hunger of religious attainments. Therefore, in order to satisfy themselves religiously, they are more involved in religious affiliations. This high involvement in fulfilling religious arrangements impacts their individual job performance which is necessary to explore. Research Methodology: Data was gathered from the public sector university employees of Pakistan. Out of 900 distributed questionnaires, 520 were received with response rate of 73.65%. Regression analysis is performed in order to determine the association between level of education and individual performance. Further, in order to determine the mediating role of religious affiliations, the Barren and Kerry (1984) model is applied. Findings: Using questionnaire survey the results of the study showed that level of education significantly impacts the religious affiliations in positive manner and high religious affiliations increase the individual job performance. The results at this point indicated that an unobservable indicator, like propensity for logical rationale induces individuals for higher education and ultimately high religious affiliations. Recommendations: Based upon the results, it is recommended that religiosity and ethical values in management set up are need to be escorted by public reforms to let the identification of employees to their work values and their obligation to the performance of work-related tasks. Originality: This study would be a clear contribution in the field of human behavior towards making alternative arrangements in order to fulfill religious obligations and at the same time identifying the mediating role of religious attainments in determining their individual performance.

Impact of Low Cost Teaching Material on Creativity, Achievement and Attitude Towards Chemistry at Secondary Level in Khyber Pakhtunkhwa

This study aimed to investigate the Impact of low Cost Teaching Material on Creativity, Achievement and Attitude towards Chemistry at Secondary level in Khyber. A related purpose was to investigate the gender differences in creativity, achievement and attitude towards Chemistry. The research strategy adopted in this study was mixed methods; both quantitative and qualitative research methods were utilized. As triangulation are the key characteristics of mixed method. Therefore, triangulation was in this research as well. To assess creativity of the students a test of creativity was developed. The test was comprised of five components, i.e. sensibility to problem, fluency, flexibility, originality and elaboration & redefinition. The reliability of the creativity test of science (Chemistry) was established using the test - retest method. The test-retest took 25 days. The test-retest reliability coefficient was found to be 0.87. Achievement test was prepared according to the style of routine exam of the schools / colleges. The objective of this particular study is to investigate the impact of teaching science through low cost materials on students’ achievement. Therefore, on the bases of cognitive domains, (Knowledge, Comprehension, Application, Analysis, Synthesis and Evaluation) of Bloom’s Taxonomy of educational objectives the researcher developed achievement tests (pre-tests and post-test) from the contents and activities of 12 grad of the Khyber Pakhtunkhwa Textbook of Chemistry. There were 29 items and every item had four options. The test was shown to practicing teachers in 15 schools and their comments were sought. And necessary changes were made in the questions and test as a whole. The test was also piloted to 80 students and item analysis was carried out. Similarly, a modified versiob of Test of Science-Related Attitude (TOSRA) was used to measure students’ attitude towards chemistry. The test was purely constructed on Likert scale to measure the students’ attitude towards chemistry on the factors such as Behavior tendency to learn chemistry, Liking for chemistry laborastory work , Liking for chemistry theory lesson , Evaluation belief about chemistry, Leisure interest in science andEnjoyment of chemistry .Besides these tests two interviews (one for teachers and one for students) were also conducted for data collection. Purposive sampling technique was adopted for selection of students for interviews. The selection of the students was non-random. The researcher selected nine students for structured interview. Three students from high achiever group three students from average and three from lower achieving groups. The selection was on the bases of achievement in their creativity test, pre-test and post-test and on the researcher observation Likert check list. There were some threats (extraneous variables) which could affect the experimental study. The researcher controlled these threats, because any uncontrolled extraneous variable could affect the performance on the depending variable to the validity of an experiment.During pilot study the researcher came to know that what type of internal and external validity could be controlled. On the bases of threats the researcher selected an institution and sample where all the threats were controlled. The most important internal threats were mortality, instrumentation, testing and differential selection of participants faced during pilot study. Due to mortality, the reduction in the number of participants occurred over time as individuals dropped out for different reasons. 26 out of 80 participants dropped out. The researcher, in broad study, overcame the mortality of groups by obtaining demographic information about the participants before the start of the study. Quantitative data was analyzed using t-test through Statistical Package for Social Sciences (SPSS). Similarly qualitative data from interviews was analyzed using qualitative data analysis techniques. Main findings of the study indicated that students in experimental group, taught through activity with low cost materials, showed significantly greater creativity, achievement and attitude towards chemistry than the students in control group. This study further showed that female students, both in experimental and control groups, outperformed their male counterparts. This shows that teaching through activities with low cost material produced better results in the terms of creativity, achievement and attitude towards chemistry.