The crude oil degradability and plasmid profile of autotrophic nitrifying bacteria, Nitrosomonas and Nitrobacter species, isolated from mangrove sediment in the Niger Delta of Nigeria were studied. The effects of temperature, pH and optical density on the utilization of different carbon sources by the bacteria were also investigated. Results showed that nitrifying bacteria could utilize kerosene, diesel oil, jet fuel and engine oil as carbon sources. None utilized hexane and xylene but moderate growth was observed in benzene, phenol and toluene. However, their ability to utilized crude oil varied both in rates of utilization and in growth profiles. Mixed culture of the isolates degrades 52 % of crude oil introduced into the medium followed by Nitrosomonas sp. with 40 % degradation. The least was Nitrobacter sp. with 20 % degradation. The ability of the autotrophs to degrade crude oil was found to be plasmid-mediated through curing experiment and electrophoresis. The size of the plasmid involved was estimated to be 23 kb. The high crude oil utilization of the mixed culture implies that nitrifying bacteria isolated from contaminated ecosystem are excellent crude oil degraders and can be harnessed for bioremediation purposes.
Polycyclic aromatic hydrocarbon (PAH)–degrading bacteria were isolated from aviation fuel contaminated soil at Inua Eyet Ikot in Ibeno, Nigeria. PAH-degrading bacteria in the contaminated soil were isolated by enrichment culture technique. Isolates with high PAH degrading potential characterized by their extensive growth on PAH-supplemented minimal salt medium were screened for their naphthalene, phenanthrene and chrysene degradability. The screening medium which contained selected PAHs as the sole source of carbon and energy showed that Micrococcus varians AFS-2, Pseudomonas putida AFS-3 and Alcaligenes faecalis AFS-5 exhibited a concentration–dependent growth in all the PAH–compounds tested. There were visible changes in the color of growth medium suggesting the production of different metabolites. Their acclimation to different PAH substrates was also evident as A. faecalis AFS-5 isolated from chrysene grew well on other less complex aromatic compounds. The isolate exhibited best growth (0.44 OD 600 ) when exposed to 10 ppm of chrysene for 5 days and could utilize up to 90 ppm of chrysene. This isolate and others with strong PAH-degrading potentials are recommended for bioremediation of PAHs in aviation fuel-contaminated sites in the tropics.
The effect of crude oil on the growth of legumes (Calopogonium muconoides and Centrosema pubescens) and fate of nitrogen-fixing bacteria in wetland ultisol was investigated using standard cultural techniques. The results revealed observable effects of oil on soil physico-chemistry, plant growth and nodulation as well as on densities of heterotrophic, hydrocarbonoclastic and nitrogen fixing bacteria. The effects however varied with different levels (0.5%, 1%, 5%, 10%, 15% and 20%) of pollution. Ammonium and nitrate levels were high in the unpolluted soil but decreased with increase in pollution levels. Nitrite was not detected in contaminated soil probably due to the reduction in numbers of nitrogen fixers, from 5.26 ± 0.23 × l06cfu/g in unpolluted soil to 9.0 ± 0.12 × 105 and 2.2 ± 0.08 × l05 cfu/g in soils with 5% and 20% levels of pollution respectively. The contaminated soil also exhibited gross reduction in the nodulation of legumes. A range of 13–57 nodules was observed in legumes from polluted soil against 476 nodules recorded for plants cultured on unpolluted soil. The heterogeneity of the microbial loads between oil-polluted and unpolluted soil were statistically significant (p < 0.05, ANOVA). Positive significant relationships were observed between the levels of hydrocarbons and the densities of heterotrophic bacteria (r = 0.91) and that of hydrocarbon utilizing bacteria (r = 0.86). On the other hand, relationships between the densities of nitrogen fixing bacteria and total hydrocarbons content was negative (r = −0.30) while positive relationships were recorded between the densities of different microbial groups and treatment periods except at 15% and 20% pollution levels. The LSD tests revealed highly significant differences (p < 0.001) in the physiological groups of soil microorganisms at all levels of pollution. The results imply that crude oil seriously affects rhizosphere microbial growth in legumes. Among the bacterial species isolated, Clostridium pasteurianum, Bacillus polymyxa and Pseudomonas aeruginosa exhibited greater ability to degrade hydrocarbons than Azotobacter sp, Klebsiella pneumoniae and Derxia gummusa while Nitrosomonas and Nitrobacter had the least degradability. A continuous monitoring of the environment is advocated to prevent extinction of nitrogen-fixing bacteria and total loss of soil fertility attributable to petroleum hydrocarbon contamination in the Niger Delta ultisol.
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