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Home > Ecology, Diversity and Metabolites Analysis of Extremophiles

Ecology, Diversity and Metabolites Analysis of Extremophiles

Thesis Info

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Author

Fariq, Anila

Program

PhD

Institute

Fatima Jinnah Women University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Environmental Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/11871/1/Anila%20Fariq%20envir%20sci%202019%20fjwu%20rwp%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676724642135

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Our earth is naturally bestowed with extreme ecological niches which rarely support any life form to exist there. Microbes harboring such habitats are unusual and exhibit specific characters to cope with stress conditions. Present research is conducted to study the diversity, ecology and metabolites of such microbes i.e. extremophiles. Water, soil and sediment samples were collected from selected extreme sites i.e. salt mine, acid seep, and hot water spring. Total DNA was extracted from soil and sediment samples for metagenome sequencing. Microbial diversity was investigated by barcoded amplicon sequencing of metagenome. Sequencing output files were submitted in the National Centre for Biotechnology Information (NCBI) database under the accession numbers SAMN08026743, SAMN08026744, SAMN08398365, and SAMN08398366 for saline, hot spring, and acid seep samples respectively. Metagenomic analysis revealed that microbial communities were dominated by Proteobacteria in all samples. Other than Proteobacteria, Firmicutes constituted the most abundant population in hot spring, Euryarchaeota were abundant in saline soil and Cyanobacteria dominated the acid seep sample. Owing to the complexities of extreme sites, some unidentified sequences were also found in samples. For isolation and characterization of extremophiles, cultivation based method was used. Eleven halophilic bacteria i.e. MB588, MB589, MB590, MB591, MB592, MB593, MB594, MB595, MB596, MB597, and MB598 were isolated from saline soil samples on LB medium with 4M NaCl concentration. Isolates were characterized morphologically, biochemically and physiologically. On the basis of 16S rDNA gene analysis, MB590, MB591, MB593, MB594, MB595, and MB596 were identified as Halomonas elongata. Whereas, MB588, MB589, MB592, MB597 and MB598 were identified as Halobacillus karajiensis, Alkalibacillus almallahensis, Aquisalibacillus elongatus, Salinicoccus sesuvii and Halomonas aquamarina respectively. Most of the strains exhibited multiple heavy metal and antibiotic resistance. Multiple antibiotics and heavy metal resistance indices i.e. MAR and MHR, revealed Halomonas elongata MB593 as a most resistant strain. Strong association among heavy metal resistance and antibiotic tolerance was observed in halophilic bacteria. No significant extremozyme production was observed in halophilic bacteria. However, biosurfactant production was indicated in nine halophilic strains.Most of the strains significantly reduced surface tension up to 35 dynes/cm and explicated emulsification activities of different hydrocarbons i.e. hexane, canola, xylene and petrol. Moreover, all the strains exhibited xviii oil displacement. Structural characterization of biosurfactants revealed mixture of glycolipids. Three bacterial strains i.e. MB592, MB597 and MB598 exhibited pigment production. Characterization of pigments using different analytical techniques i.e. UV spectroscopy, IR spectroscopy, and HPLC analysis revealed bacterioruberin carotenoids. Methanolic extracts of pigments showed strong antibacterial and antioxidant activities. From hot spring sample, seven hyper-thermophilic bacteria were isolated at 80˚C. Isolates were characterized morphologically, biochemically and physiologically. On the basis of 16S rDNA sequence analysis, isolates were identified as Geobacillus jurassicus MB599, Geobacillus jurassicus MB605, Geobacillus sp. MB601, Geobacillus sp. MB604, Geobacillus stearothermophilus MB600, Geobacillus thermocatenulatus MB602 and Geobacillus thermoparaffinivorans MB603. All the strains exhibited significant protease and amylase production. From acid seep sample, a gram negative, iron(II) oxidizing, acidophilic bacterium IO2C was isolated at pH 2 and 25˚C. On the basis of 16S rDNA analysis, it was identified as Acidithiobacillus ferrooxidans. Strain produced biominerals by oxidation of iron substrates like iron sulfate and iron ammonium sulfate. SEM, XRD and FTIR analysis of bio-minerals indicated schwertmannite and jarosite. Whole genome sequence analysis of Acidithiobacillus ferrooxidans IO-2C revealed 58.73% of G+C content, 2,716,894 base pairs in 23 contigs and 2,927 protein coding sequences. In addition to this, it contained functional genes for iron and sulfur metabolism, multidrug resistance, and heavy metal resistance. Present study provided a comprehensive examination of microbial diversity of extremophiles by consolidating culture dependent and independent techniques to get more realistic portrayal of microbial communities. This study also identified bacterial metabolites and enzymes which will provide industry with more cost-effective biomolecules for specific purposes. Future in depth studies on genomics and metabolomics of extremophilic bacterial strains identified in this study will provide insight to their adaptive mechanisms and their potential roles in bioremediation of toxic pollutants, food processing, and biomedicine.
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یہ ناداں گر گئے سجدے میںجب وقت قیام آیا

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نحمدہ ونصلی علی رسولہ الکریم امّا بعد فاعوذ بااللہ من الشیطن الرجیم
بسم اللہ الرحمن الرحیم
معزز اسا تذہ کرام اور میرے ہم مکتب ساتھیو!آج مجھے جس موضوع پر اظہار خیال کرنا ہے وہ ہے: ’’یہ ناداں گر گئے سجدے میں جب وقت قیام آیا‘‘
صدرِذی وقار!
ہر کام کا وقت مقرر ہے ، چمنستان میں گل نرگس کی شگفتگی کا ایک وقت ہے، نکہت باد بہاری کا ایک وقت ہے، کھیت اورکھلیان کے کشتِ زعفران بننے کا وقت ہے، طائر غور وفکر کی پرواز کا ایک وقت ہے، طفلانِ خود معاملہ کے عالم شباب میں پہنچنے کا ایک وقت ہے۔
صدرِمحترم!
انسان اشرف المخلوقات ہے ،شرف و بزرگی کا تاج اس کے سر سجا ہوا ہے، مجدی وسروری کی خلعتِ فاخرہ اس نے زیب تن کی ہوئی ہے ،عقل وشعور کی دولت سے مالا مال ہے ، مسجود الملائکہ کے خطاب سے نوازا گیا ہے۔ لیکن اسے بھی مسند رفیعہ کا صد رِنشیں بننے میں ایک وقت درکار ہے اس کوبھی منازل اپنے وقت پر ملتی ہیں۔
جنابِ صدر!
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صدرِمحترم!
اگر کوئی جتنا بھی فطین ہے، ذہانت و فطانت کے میدان کا شاہسوار ہے، عقل وخرداس کے گھر کی لونڈی ہے، شعور وآگہی کی فاختاؤں نے اس کے گھر میں موجو دشجر سایہ دار پر اپنا آشیانہ بنا رکھا ہو، اس کی عقل...

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Prevalence of Nasal Carriage of Methicillin-Resistant Staphylococcus Aureus in Healthcare Workers at Aga Khan University Hospital, Nairobi

Objective: To establish the prevalence of nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) in healthcare workers (HCW) at Aga Khan University Hospital, Nairobi (AKUHN). Background: Healthcare workers are a major reservoir of MRSA and may play a role in the spread of nosocomial MRSA infections. MRSA can cause significant morbidity and mortality. There is both molecular and epidemiological evidence linking HCW to MRSA outbreaks. Method: A cross sectional study conducted between July and December 2010. Nasal swabs were taken from 246 randomly selected HCW from various departments.MRSA was identified using both phenotypic and genotypic methods. Antibiotic susceptibility testing of all Staphylococcus aureus isolates was done using the broth dilution MIC technique. Results: The prevalence of nasal carriage of MRSA was 0% (95% CI:0.0%-1.5%) while that of methicillin sensitive Staphylococcus aureus (MSSA) was 18.3% (95% CI: 14.0%-23.6%). Carriage of MSSA was significantly higher in males (p=0.001, OR 5.74:2.09-15.77). Carriage was highest among HCW in the intensive care/high dependency units at 37%. Resistance to more than one class of antibiotics was seen in 37.7% of isolates. Prevalence of Panton Valentine Leukocidin (PVL) was 24.4%. Conclusion: The absence of nasal carriage of MRSA by HCW working in AKUHN suggests that they are unlikely to be the source of infections caused by MRSA in patients attending AKUHN. Screening of HCW for nasal carriage of MRSA at AKUHN should be limited to an outbreak setting. The presence of the exotoxin PVL as well as the presence of multidrug resistant strains of Staphylococcus aureus shows that HCW can still be a potential source of infections caused by MSSA.