1. EHIOSUN K.I and EZEORJIKA C.S EDO UNIVERSITY, IYAMHO
NSBMB 2017 NATIONAL CONFERENCE
University of Uyo, Akwa Ibom State
Comparative Study on Crude oil Degradation Efficiency and Peroxidase Production Dynamics by Alcaligenes eutrophs
EHIOSUN K.I and EZEORJIKA C.S
EDO UNIVERSITY, IYAMHO

2. INTRODUCTION
Crude oil pollution is a global menace that has caused critical environmental and health defects especially in oil dependent countries (Obayori et al., 2009, Muthuswamy et al., 2008).
The disadvantages associated with conventional treatment of crude oil pollution have led to the development of an alternative, more economical and reliable biological approach. This makes the search for crude oil degrading microbes very crucial (Olajuyigbe and Ehiosun, 2016).

3. INTRODUCTION CONT’D
Studies have shown that certain species of bacteria called hydrocarbon utilizing bacteria (HUB) could degrade crude oil hydrocarbons in the presence of carbon, hydrogen, oxygen and nitrogen to stimulate their growth and multiplication (Nester et al, 1998; Agunwamba et al, 2004).
Extracellular and intracellular degradation of organic pollutants by microbes is an oxidative process catalyzed by peroxidases and oxygenases (Van-Beilen and Funhoff, 2007).

4. Poly aromatic hydrocarbons
Introduction Cont’d
65%
Enzymes
Poly aromatic hydrocarbons
Peroxidase
(Baborová et al., 2006; Eibes et al., 2006)

5. Aim
The present study became necessary to investigate the crude oil biodegradation activity of selected bacterial isolate and evaluate the correlation between degradation efficiency and peroxidase production.

6. Objectives of Research
The objectives of this research were to:
Isolate and identify microbial isolates from crude oil contaminated soil capable of crude oil degradation.
Carry out biodegradation of varying concentration of crude oil with the selected isolate.
Monitor microbial growth, residual crude oil and peroxidase production.
Evaluate the correlation between degradation efficiency and peroxidase production.

7. Materials and Methods Soil collection (Aslim et al., 2002)
Microbial isolation, characterisation and Identification (Bergey’s Manual of Systematic Bacteriology, 2009)
Screening bacterial isolates on glucose and crude oil media
Biodegradation of varying crude oil concentration (Muthuswamy et al., 2008)

8. Materials and Methods Cont’d
Estimation of residual crude oil concentration and degradation efficiency (Odu et al., 1985)
Determination of peroxidase production (Hunter et al., 2003)

9. RESULTS AND DISCUSSION

10. Table1: Morphological and biochemical characterization of bacteria from 20 cm soil depth
Morphological Test
Biochemical Test
Microbial
identity
Colour
Gram reaction
Cellular morphology
Catalase test
Oxidase Test
Indole test
Motility test
Methyl red test
Voges-Prokauer Test
Citrate utilization
Urease activity
Starch hydrolysis
Gelatin hydrolysis
Casein hydrolysis
Spore test
NO3 reduction
Glucose
Sucrose
Arabinose
Maltose
Mannitol
Xylose
Galactose
Raffinose
Saucin
Sorbitol
Lactose
Cream +
Rods -
Corynebacterium striatum
Bacillus polymyxa
Alcaligenes eutrophs
Bacillus coagulans

11. Screening of Bacteria Isolates
Figure 1: Growth profile of bacterial isolates on crude oil and glucose based media (a) Corynebacterium striatum (b) Bacillus polymyxa (c) Alcaligenes eutroph (d) Bacillus coagulans

12. Crude oil Biodegradation Study
Figure 2: Growth profile and residual crude oil of Alcaligenes eutrophs in (a) 10 g/L (b) 30 g/L (c) 50 g/L (d) 75 g/L crude oil based medium

13. Crude oil Biodegradation Study
Figure 3: Degradation efficiency and peroxidase production by Alcaligenes eutrophs in (a) 10 g/L, (b) 30 g/L, (c) 50 g/L, (d) 75 g/L crude oil based medium.

14. CONCLUSION
The remarkable crude oil degradation properties exhibited by Alcaligenes eutrophs in this study had shown how tolerant and efficient the isolate is toward varying concentration of crude oil. The high activity of peroxidase coupled with the removal of crude oil is indicative of a relationship between the enzyme’s activity and biodegradation yield. These properties suggest that the microbe under study is a good bio-resource for crude oil bioremediation.

15. REFERENCE
Agunwamba, J.C, Ezeogu, L.I and Chukwu, I. (2004). Effects of nutrient, crude oil, and Pseudomonas concentrations I: Bacteria Growth. Int. J. of Envi.Issues, pp
Aslim, B., Z. N. Yuksekdag, and Y. Beyatli (2002). Determination of PHB growth quantities of certain Bacillus species isolated from soil. Turk. Electron. J. Biotechnol. (Special Issue):24–32.
Hunter C. L., Maurus R., Mauk M. R., Lee H., Raven E. L., Tong H., Nguyen N., Smith M., Brayer G. D., and Mauk A. G. (2003). Introduction and characterization of a functionally linked metal ion binding site at the exposed heme edge of myoglobin. PNAS 100(7): 3647–3652.

16. REFERENCE
Muthuswamy, S., R. B. Arthur, B. Sang-Ho, and Y. Sei-Eok (2008). Biodegradation of crude oil by individual bacterial strains and a mixed bacterial consortium isolated from hydrocarbon contaminated areas. Clean 36:92–96.
Nester, E.W., Roberts, C.E., Pearsall, N.N., Anderson, D.G. and Nester, M.T (1998). Microbiology: A human perspective. 2nd ed. New York McGraw Hill.
Obayori, O. S., M. O. Ilori, S. A. Adebusoye, G. O. Oyetibo, A. E. Omotayo, and O. O. Amund (2009). Degradation of hydrocarbons and biosurfactant production by Pseudomonas sp. strain LP1. World J. Microbiol. Biotechnol. 25:1615–1623.

17. REFERENCE
Odu, C. T. I, Esuruoso O. F., Nwoboshi L.C. and Ogunwale J.A (1985).  Environmental Study of the Nig. Agip Oil Company, Operational Area.  Soil and Fresh Water Vegetation.  Union Graft publs.  Milan pp. 22 – 25.
Olajuyigbe Folasade M. and Ehiosun Kevin I. (2016). Assessment of crude oil degradation efficiency of newly isolated actinobacteria reveals untapped bioremediation potentials. Bioremediation Journal, 20(2):
Van-Beilen J.B and Funhoff E.G (2007). Alkane hydroxylases involved in microbial alkane degradation. Appl. Microbiol. Biotechnol. 74:13-21.

18. APPRECIATION EDO UNIVERSITY, IYAMHO
FEDERAL UNIVERSITY OF TECHNOLOGY, AKURE

19. THANK YOU