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Assessment of population dynamics of biodegrading consortia in bonny light crude oil polluted agricultural soil using molecular and culture-based technique
Population dynamics of biodegradable consortia in crude oil polluted soil was undertaken. Culture-based and molecular
biology technique was employed. The result of the total heterotrophic bacteria count (THC) in cfu/ml showed higher
increase in polluted soil on comparison to control sample irrespective of the medium used. Diverse microorganisms were
isolated and identified using culture based approach. Bacterial isolates obtained include Acinetobacter, Pseudomonas,
Mycobacterium, Proteus mirabilis, Streptococcus, Actinomyces sp, Micrococcus luteus, Bacillus sp, Corynebacterium sp,
Lactobacillus sp, E. coli, Providencia, Citrobacter freundii, Enterobacter, Staphylococcus and Alcaligens. Fungal isolates obtained
also include Rhizopus, Penicillium, Fusarium, Aspergillus niger, Aspergillus flavus, Aspergillus nidulans, Aspergillus fumigatus,
Eurothium, Cladosporium, Brettanomyces, Candida albican and Murtierella. Scoring on the persistence of isolates in polluted
soil showed that Micrococcus, Bacillus sp, E. coli and Staphylococcus sp had 33.3% persistence followed by Lactobacillus sp,
Providencia sp, Citrobacter and Bacillus sp (subspecies) which recorded 66.6% persistence whereas others persisted throughout
the study. Molecular identification of organisms using specific primers as C230 had bands depicting similarity of about 90%
to catechol 2, 3-dioxygenase gene with significant alignment for Bacillus megaterium, Bacillus cereus, Micrococcus luteus and
Lactobacillus acidophilus. ACT and OM primer produced 100% significant alignment for Corynebacterium, Actinomyces,
Candidatus, Streptococcus, Streptomyces and Enterobacter cloacca, Streptococcus salivarius, Yerisinia sp respectively. Random
amplified polymorphic DNA (RAPD) assay using specific oligonucleotide primer OP sequence of 5ΓΆΒ?Β? GTGACGTAGG 3ΓΆΒ?Β? was
employed to generate several distinct DNA product which primed at multiple locations throughout the genome of test isolates.
Spectrum of amplified product was evident with B2, B16, B17 and B30 isolates. Although distinct tree obtained using PyELph
1.4 software elucidated that B16 and B17 which are gram positive rods are not closely related but the position of band 3 and
4 of both isolate indicates that they may have similar traits of negligible identity. Statistical analysis however showed that the
volume of crude oil in relation to weeks of treatment have significant interaction effects on the total heterotrophic count.
Biography
Ogbulie Toochukwu Ekwutosi has completed her PhD from Federal University of Technology Owerri, Nigeria. She has also obtained additional qualification as Registered Environmental Scientist by NREP, USA in 2008. She is a Senior Lecturer in the Department of Biotechnology, FUTO, Nigeria. She has published more than 30 papers in reputed journals and has been serving as an Editorial Board Member of Analele Universitatii din Oradea-Fascicula Biologie, Romania and Romanian Biotechnology Letters. She is a Member of societal bodies such as Society for Applied Microbiology UK (SFAM), Graduate Women in Science GWIS USA, Organization for Women in Science for the Developing World (OWSD), Biotechnology Society of Nigeria & Nigerian Society for Microbiology, Nigerian Environmental Society and National Registry of Environmental Professionals, USA.