Biodegradation of Total Petroleum Hydrocarbon by Aerobic Heterotrophic Bacteria Isolated from Crude Oil Contaminated Brackish Waters of Bodo Creek

Abstract

Bodo creek is located in Gokana Local Government Area of Rivers state and is characterised by brackish water which is heavily contaminated with crude oil. Water samples collected from the creek were taken to Environmental Microbiology Laboratory of University of Portharcourt for isolation of aerobic heterotrophic bacteria using serial dilution spread plate technique. Isolation of morphologically distinct pure cultures was done and the isolates obtained were identified molecularly on the basis of 16S rRNA gene sequence analysis. 16S rRNA sequencing for identification of the isolates generated sequences ranging from 500 bp to 1700 bp and a 250 bp size PCR amplified fragment. The nucleotide sequence of the 16S rRNA gene was compared with published 16S rRNA sequences using BLAST search at the data base of National Centre for Biotechnology Information (NCBI). Biodegradation of petroleum hydrocarbon test on the isolates was conducted using Bony light crude oil obtained from Shell Petroleum Development Company and monitored using GC-FID (Agilent 6890 model) for 49 days. The initial quantity of TPH was 24091 and 6706 mg/l on day 0 for aerobic heterotrophic treatment and crude oil contaminated control and decreased progressively to 212.8 and 1174 mg/l respectively on day 49 indicating biodegradation in the treatment and control. Loss of TPH was statistically significant using one way analysis of variance (ANOVA) p<0.05 with time. Model for predicting TPH loss was developed and rate of biodegradation of the isolates determined using Trend line method of Microsoft Excel, 2010. The growth of bacteria cells increased progressively with decrease in TPH implying that the bacteria were responsible for biodegradation. Further application of bioremediation strategies in Bodo creek for biostimulation of the crude oil biodegrading bacteria for reclamation and restoration of an efficient ecosystem structure and function is suggested.