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Profiling of Indigenous Bacterial Strains for the Remediation of Industrial Effluents

Thesis Info

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Author

Fozia Aslam

Program

PhD

Institute

Fatima Jinnah Women University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2016

Thesis Completion Status

Completed

Subject

Environmental Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/14192/1/Fozia_Aslam_Envir_Science_HSR_2017_FJWU_Rwp_07.02.2018%20%28revised%29.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676724948915

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Industrial revolution has played a positive role in the economic growth and development but environment is affected negatively. This has led to the contamination of environment with toxic chemicals. Biodegradation of these harmful pollutants using microbes is strongly recommended as this is more efficient than other costly physicochemical processes. Current study was aimed to isolate indigenous bacteria capable of biodegrading toxic chemicals. For this purpose effluent samples were collected from Hattar Industrial Estate, Haripur, Pakistan and characterized. Initially, forty eight distinct bacterial isolates were obtained by plating on mixed metal nutrient agar medium supplemented with 200μg/ml each of chromium, lead, nickel salts. Further screening on solid agar medium both in rich and minimal medium (M9 and BH) supplemented with 12 different metal salts (Cr, Cu, Cd, As, Ag, Hg, Zn, Fe, Mn, Ni and Co) was done. Representatives of different classes of aromatic compounds including polyaromatic hydrocarbons phenanthrene (500-6000 μg/ml), biphenyl and naphthalene (1000-4000 μg/ml); petroleum hydrocarbons (1-3%) benzene, toluene, xylene; aromatic amines (1mM) aniline, chloroaniline; phenolic compounds (50-200μg/ml) phenol, p-nitrophenol, pentachlorophenol and azo dyes (50-100μg/ml) methyl orange, methyl yellow, methyl red, erichrome black T and congo red were used to determine the degradation potential of isolates. Results revealed that these bacteria had multiple heavy metal resistances. They were potentially able to utilize multiple hydrocarbons and azo dyes as carbon/ energy sources after degradation. Based on these tolerance/utilization profiles sixteen morphologically, physiologically and biochemically different isolates were chosen for metal remediation/ organic pollutant utilization/ degradation studies. Quantitative and qualitative investigation of transformation ability of these sixteen isolates for phenanthrene (1000mg/l), aniline (1mM), p-nitrophenol (50mg/l), pentachlorophenol (100mg/l) and azo dyes (50mg/l) congo red, methyl red was achieved by high performance liquid chromatography (HPLC), UV-visible spectroscopy and Fourier transform infrared spectroscopy (FTIR). Five potent strains were identified as Stenotrophomonas sp. MB339 (Acc No. KP723528), Staphylococcus sp. MB371 (Acc No. KP723530), Bacillus sp. MB366 (Acc No. KP723529), Klebsiella pneumoniae MB361 (Acc No. KP723532), Klebsiella pneumoniae MB373 (Acc No. KP723531) by 16S rRNA gene sequencing. Among metal remediating strains, Staphylococcus MB371, Stenotrophomonas MB339, Klebsiella pneumoniae MB361 showed maximum accumulation of Pb, Cr and Ni (i.e. 75.33, 34.07, 54.98%), respectively at 37°C. In addition, these strains efficiently reduced more than 70% of 200mg/l Cr (VI) to Cr (III) and >90% of phenanthrene, p-nitrophenol, pentachlorophenol at different pHs (5-9) and temperature (25-45°C). In case of aniline up to 91% degradation was achieved by Bacillus MB366, Stenotrophomonas MB339, Klebsiella pneumoniae MB373 at pH 7, 37°C. While, Staphylococcus sp. MB371, Klebsiella pneumoniae MB361 were unable to use aniline at pHs 5, 6. Decolorization of methyl red (>75%) was achieved by all five strains within pH range 5-9 and 30-37°C. However, less or even no decolorization was observed at 45°C especially by Stenotrophomonas MB339. Complete genome sequence analysis of these five strains was done using Ilumina Miseq and annotated with IMG ER pipeline. Genomic features of strains were different from one another in many attributes like size, total number of genes, protein coding genes, rRNA genes and G+C content. Functional annotations depicted that these strains were highly diverse in their metabolism. Heavy metal resistance genes organized in operons for arsenic, copper, mercury were identified in Stenotrophomonas MB339, for arsenic in Staphylococcus MB371 and Bacillus MB366. While genes for metal (Cr, As) reductase, efflux pump and other membrane proteins (Co, Cd, Fe, Zn, Mn) were annotated in genomes of Klebsiella quasipneumoniae subsp. similipneumoniae MB373, Stenotrophomonas MB339, Staphylococcus MB371 and Bacillus MB366. Genome annotations indicated that these genomes have variety of metabolic pathways for xenobiotics degradation. Aromatic ring hydroxylating dioxygenase genes responsible for degradation of PNP, PCP, phenanthrene. While genes for FMN-dependent, NADH-azo reductase might be supportive for methyl red decolorization. For nitroaromatic compounds, nitroreductase genes were found in Klebsiella pneumoniae MB361, Stenotrophomonas MB339 and Bacillus MB366. Indigenous bacteria isolated from industrial effluents play a significant role to detoxify metals and transform variety of chemicals present in contaminated environment. Present study revealed the molecular mechanisms behind adaptability of these biotopes to changing environmental conditions. This detailed research comprising of analytical and molecular approaches, highlight the biodegradation potential of strains which paves way for future bioprospecting of the potential enzymes vital for biodegradation perspective.
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41:01
a. Ha. Mim.

41:02
a. This is a Revelation from Allah - The One and Only God of everyone and everything.
b. The Immensely Merciful to all, The Infinitely Compassionate to everyone.

41:03
a. This is a Book whose Messages are clearly explained and well spelled-out -
b. Qur’an in Arabic -
c. for a people who understand.

41:04
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41:05
a. And they say:
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41:06
a. Tell them O The Prophet:
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