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Screening of polycyclic aromatic hydrocarbon degrading bacterial isolates from oil refinery wastewater and detection of conjugative plasmids in polycyclic aromatic hydrocarbon tolerant and multi-metal resistant bacteria.

ABSTRACT: Wastewater were collected from the effluent channel in the vicinity of Mathura oil refinery, U.P. (India) and analysed for physicochemical characteristics, heavy metals as well as organic compounds including PAHs. The interaction of PAHs and heavy metals with various group of microorganisms revealed the viable count of aerobic heterotrophs, asymbiotic nitrogen fixers, actinomycetes and fungi were found to be 2.38 × 106, 1.89 × 104, 2.20 × 104 CFU/mL and 8.76 × 103 CFU/mL respectively. We have selected and screened 50 bacterial isolates for their resistance/tolerance to heavy metal and PAHs. Out of 25 multi-metal resistant isolates, 6 were able to tolerate PAHs at the concentration of 5000 ?g/mL (50?g/disc) to naphthalene, anthracene, phenanthrene and pyrene. The PAH degradation efficiency of the isolates was assessed using spectrophotometer with 100 ?g/mL of phenanthrene and observed different degree of degradation ranging from 34-66% after 96 h of incubation. One of the bacterial isolates KWB3 (identified as Enterobacter ludwigii by 16S rDNA sequencing) exhibited maximum degradation efficiency (66%) was further tested for phenanthrene degrading ability in the presence and absence of a co-substrate (glucose) in a mineral salt medium; and a number of metabolites were produced and detected by GC-MS which revealed the presence of benzocoumarin, phthalic acid, catechol and several low molecular weight compounds. The DNA derived from multi-metal and PAHs tolerant bacteria were PCR amplified using Inc specific primers and positive PCR products were obtained with oriT and trfA2 of the IncP group; indicates that these bacteria have gene-mobilizing capacity.

SUBMITTER: Khatoon K

PROVIDER: S-EPMC6838955 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Screening of polycyclic aromatic hydrocarbon degrading bacterial isolates from oil refinery wastewater and detection of conjugative plasmids in polycyclic aromatic hydrocarbon tolerant and multi-metal resistant bacteria.

Khatoon Khalida K Malik Abdul A

Heliyon 20191001 10

Wastewater were collected from the effluent channel in the vicinity of Mathura oil refinery, U.P. (India) and analysed for physicochemical characteristics, heavy metals as well as organic compounds including PAHs. The interaction of PAHs and heavy metals with various group of microorganisms revealed the viable count of aerobic heterotrophs, asymbiotic nitrogen fixers, actinomycetes and fungi were found to be 2.38 × 106, 1.89 × 104, 2.20 × 104 CFU/mL and 8.76 × 10 ...[more]

PMID: 31720477

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Project description:The characterization of native polycyclic aromatic hydrocarbon (PAH)-degrading bacteria is significant for understanding the PAH degradation process in the natural environment and developing effective remediation technologies. Most previous investigations of PAH-degrading bacteria in environmental samples employ pahAc, which encodes the ?-subunit of PAH ring-hydroxylating dioxygenase, as a functional marker gene. However, the poor phylogenetic resolution and nonspecificity of pahAc result in a misestimation of PAH-degrading bacteria. Here, we propose a PAH hydratase-aldolase-encoding gene, pahE, as a superior biomarker for PAH-degrading bacteria. Comparative phylogenetic analysis of the key enzymes involved in the upper pathway of PAH degradation indicated that pahE evolved dependently from a common ancestor. A phylogenetic tree constructed based on PahE is largely congruent with PahAc-based phylogenies, except for the dispersion of several clades of other non-PAH-degrading aromatic hydrocarbon dioxygenases present in the PahAc tree. Analysis of pure strains by PCR confirmed that pahE can specifically distinguish PAH-degrading bacteria, while pahAc cannot. Illumina sequencing of pahE and pahAc amplicons showed more genotypes and higher specificity and resolution for pahE Novel reads were also discovered among the pahE amplicons, suggesting the presence of novel PAH-degrading populations. These results suggest that pahE is a more powerful biomarker for exploring the ecological role and degradation potential of PAH-degrading bacteria in ecosystems, which is significant to the bioremediation of PAH pollution and environmental microbial ecology.IMPORTANCE PAH contamination has become a worldwide environmental issue because of the potential toxic effects on natural ecosystems and human health. Biotransformation and biodegradation are considered the main natural elimination forms of PAHs from contaminated sites. Therefore, the knowledge of the degradation potential of the microbial community in contaminated sites is crucial for PAH pollution bioremediation. However, the nonspecificity of pahAc as a functional marker of PAH-degrading bacteria has resulted neither in a reliable prediction of PAH degradation potential nor an accurate assessment of degradation. Here, we introduced pahE encoding the PAH hydratase-aldolase as a new and better functional marker gene of PAH-degrading bacteria. This study provides a powerful molecular tool to more effectively explore the ecological role and degradation potential of PAH-degrading bacteria in ecosystems, which is significant to the bioremediation of PAH pollution.

Project description:BackgroundPolycyclic aromatic hydrocarbons (PAH) are produced by the burning and processing of fuel oils, and have been associated with oxidant stress, insulin resistance and hypertension in adults. Few studies have examined whether adolescents are susceptible to cardiovascular effects of PAHs.ObjectiveTo study associations of PAH exposure with blood pressure (BP) and brachial artery distensibility (BAD), an early marker of arterial wall stiffness, in young boys attending three schools in Jeddah, Saudi Arabia in varying proximity to an oil refinery.MethodsAir samples collected from the three schools were analyzed for PAHs. PAH metabolites (total hydroxyphenanthrenes and 1-hydroxypyrene) were measured in urine samples from 184 adolescent males, in whom anthropometrics, heart rate, pulse pressure, brachial artery distensibility and blood pressure were measured. Descriptive, bivariate and multivariable analyses were performed to assess relationships of school location and urinary PAH metabolites with cardiovascular measures.ResultsTotal suspended matter was significantly higher (444 ± 143 μg/m(3)) at the school near the refinery compared to a school located near a ring road (395 ± 65 μg/m(3)) and a school located away from vehicle traffic (232 ± 137 μg/m(3)), as were PAHs. Systolic (0.47 S D units, p = 0.006) and diastolic (0.53 SD units, p < 0.001) BP Z-scores were highest at the school near the refinery, with a 4.36-fold increase in prehypertension (p = 0.001), controlling for confounders. No differences in pulse pressure, BAD and heart rate were noted in relationship to school location. Urinary total hydroxyphenanthrenes and 1-hydroxypyrene were not associated with cardiovascular outcomes.ConclusionsProximity to an oil refinery in Saudi Arabia is associated with prehypertension and increases in PAH and particulate matter exposures. Further study including insulin resistance measurements, better control for confounding, and longitudinal measurement is indicated.

Dataset Information

Screening of polycyclic aromatic hydrocarbon degrading bacterial isolates from oil refinery wastewater and detection of conjugative plasmids in polycyclic aromatic hydrocarbon tolerant and multi-metal resistant bacteria.

Publications

Screening of polycyclic aromatic hydrocarbon degrading bacterial isolates from oil refinery wastewater and detection of conjugative plasmids in polycyclic aromatic hydrocarbon tolerant and multi-metal resistant bacteria.

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