An Investigation of Petrol Metabolizing Bacteria Isolated from Contaminated Soil Samples Collected from Various Fuel Stations

Abstract

The present study aimed to isolate the high-efficiency petrol metabolizing thermophilic bacteria from petrol contaminated soil samples. Isolation was carried out through enrichment culture, serial dilution and pour plate methods using the petrol supplemented minimal salt media. The isolated bacteria were analyzed to document growth behavior, petrol removal efficiencies, antibiotic resistance profile, and biochemical characteristics. The 16S rRNA based phylogenetic analysis helped to reveal the identity of isolated bacterial species and construct the phylogenetic trees. Total nine bacteria were isolated, out of which three (IUBP2, IUBP3, IUBP5) were identified as Brevibacillus formosus, one (IUBP1) was found similar to Brevibacillus agri, four (IUBP7, IUBP8, IUBP13, and IUBP14) shared homology with Burkholderia lata, and one (IUBP15) with Burkholderia pyrrocinia. All the isolates were fast growing and exhibited considerable petrol degradation potential. The highest petrol removal efficiency (69.5% ± 13.44/6 days) was recorded for the strain IUBP15 at a petrol concentration of 0.1% (v/v). All bacteria studied (100%) were positive for esculinase and phosphatase. Many strains exhibited positive responses for arginine dehydrolase (22%), β-naphthylamidase (11%), β-D-glucosaminide (33%), mannitol (55%), sorbitol (66%) and inulin (88%) fermentation test. While all were sensitive to the antibiotics, some of them were found resistant against chloramphenicol and oxacillin. The remarkable biochemical characteristics and considerable petrol removal potential (40–70%) highlights utilization of the bacteria isolated for petrol bioremediation, mineralization of organophosphates, dairy and food industry, and also as biofertilizers and biocontrol agents.

Molecular and metabolic characterization of petroleum hydrocarbons degrading Bacillus cereus

“…CT10) and 48 hr ( Pseudomonas and Bacillus sp.) ( Liang et al 2018 ; Tian et al 2018 ; Muccee and Ejaz 2019 ).…”

“…Various petroleum degraders have been biochemically characterized in previous studies and found positive for the assays like catalase test, MacConkey agar test, lactose fermentation, mannitol test, urea hydrolysis test, starch hydrolysis test, citrate test, carbohydrate utilization test, esculinase test, phosphatase test, PYR, ADH, β-D-glucosaminidase test, mannitol, sorbitol and inulin fermentation tests ( Muccee and Ejaz 2019 ; Hossain et al 2022 ). So, the present study’s findings are consistent with earlier research, as B. cereus has also been found positive for ADH.…”

“…For quantitative estimation of the TPH removal efficiency of bacteria, a previously reported method involving the use of 2,6-dichlorophenolindophenol (DCPIP) as a redox indicator, was employed ( Bilen Ozyurek and Seyis Bilkay 2017 ; Marchand et al 2017 ; Muccee and Ejaz 2019 ). To perform this assay, 1% v/v of DCPIP was added to MSM containing petroleum as a carbon source.…”

Molecular and functional analysis of naphthalene‐degrading bacteria isolated from the effluents of indigenous tanneries

“…Conditions used for PCR amplification were; initial denaturation for 5 min. at 95°C, 38 cycles of initial denaturation for 40 s at 94°C, annealing for 60 s at 58°C, extension for 30 s at 72°C, and final extension for 10 min at 72°C [20]. Amplified PCR products of 1500 bp were subjected to agarose gel electrophoresis and purified using Monarch DNA Gel Extraction Kit (Cat # T1030S BioLabs) and sent for sequencing to Macrogen.…”

“…Extracellular and intracellular metabolites were extracted using a previously reported protocol [20]. Supernatant of a bacterial culture was used as a source of extracellular metabolites and mixed with absolute methanol in equimolar concentration.…”