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Mechanic workshops in residential areas are sites referenced for small-scale chronic contamination with petroleum hydrocarbons (HC) and associated pollutants like heavy metals. The main aim of this study was to isolate from a mechanic workshop soil in Lagos a novel indigenous bacterium with potential for degradation of phenanthrene, a model polyaromatic compound. Gas chromatographic analysis revealed concentrations of HC (20,055 mg kg-1), lead (156.19 mg kg-1) and zinc (202.005 mg kg-1) in excess of regulatory limits in the soil. Continuous enrichment resulted in the isolation of a bacterium strain ALSL2, which was identified by 16S rRNA gene sequencing as Bacillus thuringiensis. Strain ALSL2 showed broad specificity for a range of HCs including phenanthrene, anthracene, biphenyl, dibenzothiophene, crude oil, diesel and kerosene and also showed biosurfactant production with emulsification index 55.2 %, 65.7 % and 58.7 % on crude oil, kerosene and vegetable oil respectively. The isolate showed evidence of dioxygenase activity and utilized metabolites of aromatic hydrocarbon degradation including 1-naphthol and O-phthalate. At the end of 10 days 51.45 % of phenanthrene was degraded at the rate of 7.922 mg l−1 d−1, degradation constant 0.073 d−1, and half-life 9.864 d. The corresponding values for anthracene were 69.11 %, 9.92 mg l−1 d−1., 0.11 d−1, and 5.924 d respectively. Our findings represent a remarkable addition to available indigenous bioresources with potential for application in bioremediation, and further highlight mechanic workshop soils as veritable source of such isolate.

Abstract Background Cytochrome P450 monooxygenases (CYPs) are exciting biocatalysts that catalyzes diverse regio- and stereoselective reactions of a broad range of substrates. The cytochrome P450 genes (CYPomes) of a chronically polluted soil (3S) with history of agricultural activities were deciphered via functional annotation of putative ORFs (open reading frames) using KEGG KofamKOALA, PHMMER, the Cytochrome P450 Engineering Database CYPED v6.0, and the NCBI Batch Web CD-Search tool. Results Annotation revealed the detection of seventy-seven CYP families and eight standalone CYPs cutting across the three domains of life. The prokaryote CYPome has 72 CYP families, 93 subfamilies, and seven standalone CYPs. The phylum Proteobacteria and the genera Streptomyces, Mycobacterium, and Bacillus with 17, 16, 24, and 5 CYP families were predominant, while the domain Archaea was represented by CYP119A1. The phylum Cyanobacteria has two families, while 23 actinobacterial CYPs (other than Streptomyces and Mycobacterium) were also detected. The detected prokaryote CYPs are responsible for biodegradation of camphor, hydroxylation of monoterpene alcohols, biosynthesis of secondary metabolites, and hydroxylation of fatty acids and steroidal compounds. The eukaryote CYPome was represented by seven fungal CYPs (CYP505A3, CYP505B1, CYP51A, CYP51C, CYP55A1, CYP55A2, and CYP55A3) from Acremonium egyptiacum, Fusarium oxysporum, Aspergillus oryzae, Gibberella moniliformis, Aspergillus flavus, and Fusarium lichenicola, respectively, and CYP524A1 from the slime mold, Dictyostelium discoideum. The fungi CYPs were involved in biosynthesis of secondary metabolites, hydroxylation of fatty acids, and nitrate reduction and denitrification. Conclusions This study has established the diverse roles played by CYPs in soil, its implication for soil health and resilience, and its potentials for industrial application.

Abstract Background Soils polluted with animal charcoal from skin and hide cottage industries harbour extremely toxic and carcinogenic hydrocarbon pollutants and thus require a bio-based eco-friendly strategy for their depuration. The effects of carbon-free mineral medium (CFMM) amendment on hydrocarbon degradation and microbial community structure and function in an animal charcoal-polluted soil was monitored for 6 weeks in field moist microcosms consisting of CFMM-treated soil (FN4) and an untreated control (FN1). Hydrocarbon degradation was monitored using gas chromatography-flame ionization detector (GC-FID), and changes in microbial community structure were monitored using Kraken, while functional annotation of putative open reading frames (ORFs) was done using KEGG KofamKOALA and NCBI’s conserved domain database (CDD). Results Gas chromatographic analysis of hydrocarbon fractions revealed the removal of 84.02% and 82.38% aliphatic and 70.09% and 70.14% aromatic fractions in FN4 and FN1 microcosms in 42 days. Shotgun metagenomic analysis of the two metagenomes revealed a remarkable shift in the microbial community structure. In the FN4 metagenome, 92.97% of the population belong to the phylum Firmicutes and its dominant representative genera Anoxybacillus (64.58%), Bacillus (21.47%) and Solibacillus (2.39%). In untreated FN1 metagenome, the phyla Proteobacteria (56.12%), Actinobacteria (23.79%) and Firmicutes (11.20%), and the genera Xanthobacter (9.73%), Rhizobium (7.49%) and Corynebacterium (7.35%), were preponderant. Functional annotation of putative ORFs from the two metagenomes revealed the detection of degradation genes for aromatic hydrocarbons, benzoate, xylene, chlorocyclohexane/chlorobenzene, toluene and several others in FN1 metagenome. In the FN4 metagenome, only seven hydrocarbon degradation genes were detected. Conclusion This study revealed that though CFMM amendment slightly increases the rate of hydrocarbon degradation, it negatively impacts the structural and functional properties of the animal charcoal-polluted soil. It also revealed that intrinsic bioremediation of the polluted soil could be enhanced via addition of water and aeration.

Abstract Background The purpose of this study is to use shotgun next-generation sequencing to unravel the microbial community structure of an agricultural soil, decipher the effects of mercury contamination on the structure of the microbial community and the soil physicochemistry and heavy metals content. Results The soil physicochemistry after mercury contamination revealed a shift in soil pH from neutral (6.99 ± 0.001) to acidic (5.96 ± 0.25), a decline in moisture content to < 4 %, and a significant decrease in the concentrations of all the macronutrients and the total organic matter. Significant decrease in all the heavy metals detected in the agricultural soil was also observed in mercury inundated SL3 microcosm. Structural analysis of the metagenomes of SL1 (agricultural soil) and SL3 (mercury-contaminated agricultural soil) using Illumina shotgun next-generation sequencing revealed the loss due to mercury contamination of 54.75 % of the microbial community consisting of an archaeal domain, 11 phyla, 12 classes, 24 orders, 36 families, 59 genera, and 86 species. The dominant phylum, class, genus, and species in SL1 metagenome are Proteobacteria, Bacilli, Staphylococcus, and Sphingobacterium sp. 21; while in SL3 metagenome, Proteobacteria, Alphaproteobacteria, Singulisphaera, and Singulisphaera acidiphila were preponderant. Mercury contamination resulted in a massive upscale in the population of members of the phylum Planctomycetes and the genera Singulisphaera, Brevundimonas, Sanguibacter, Exiguobacterium, Desulfobacca, and Proteus in SL3 metagenome while it causes massive decline in the population of genera Staphylococcus and Brachybacterium. Conclusions This study revealed that mercury contamination of the agricultural soil imposed selective pressure on the members of the microbial community, which negatively impact on their population, alter soil physicochemistry, and enriched sizable numbers of members of the community that are well adapted to mercury stress. It also reveals members of microbial community hitherto not reported to be important in mercury detoxification process.

Abstract Background The purpose of this study is to decipher the diverse carbohydrate metabolism pathways in a spent engine oil-perturbed agricultural soil, enunciate the carbohydrate-active enzymes and genes involved in the process, taxonomically classify the annotated enzymes and genes, and highlight the importance of the study for ecological and biotechnological processes. Results Functional analysis of the metagenome of spent engine oil (SEO)-contaminated agricultural soil (AB1) using the Kyoto Encyclopedia of Genes and Genomes (KEGG) GhostKOALA, Cluster of Orthologous Groups (COG) of proteins, the Carbohydrate-Active Enzymes (CAZy) database, and the NCBI’s conserved domain database (CDD) revealed extensive metabolism of carbohydrates via diverse carbohydrate-active enzymes and genes. Enzymes and genes annotated for glycolysis/gluconeogenesis pathway, citric acid (TCA) cycle, pentose phosphate pathway, and pyruvate metabolism, among others, were detected, and these were not detected in the original agricultural soil (1S). Analysis of carbohydrate-active enzymes, using the CAZy database, showed 45 CAZy families with preponderance of glycoside hydrolases (GHs, 46.7%), glycosyltransferases (GTs, 24.4%), and carbohydrate-binding modules (CBMs, 15.5%). Taxonomic classification of the annotated enzymes and genes for carbohydrate metabolism using the GhostKOALA and CAZy databases revealed the predominance of the phylum Proteobacteria with the representative genera Pseudomonas (18%), Sphingobium (13.5%), and Sphingomonas (4.5%), respectively. Biotechnologically important enzymes such as xylanases, endoglucanases, α- and β-glucosidases and glycogen debranching enzymes were also retrieved from the metagenome. Conclusions This study revealed the presence of diverse carbohydrate-active enzymes and genes mediating various carbohydrate metabolism pathways in the SEO-perturbed soil metagenome. It also reveals the detection of biotechnologically important enzymes with potentials for industrial use.