Scientific Research and Essays Vol. 7(1), pp.

66-77, 9 January, 2012

Available online at http://www.academicjournals.org/SRE
DOI: 10.5897/SRE11.1146 ISSN 1992-2248 2012 Academic Journals
Biodegradation of pyrene by a newly isolated Proteus vulgaris
Nur Ceyhan
Bacterial growth on plate medium supplemented with pyrene
Carbon-free mineral medium (CFMM) of Yuting et al. (2003) and Habe et al. (2004) was
prepared in plates with some modifications. The CFMM used as the basal medium contained (per
liter of distilled water) (NH4)2SO4 2.0 g, NaH2PO4 0.5 g, K2HPO4 0.5 g, MgSO47H2O 0.2 g,
CaCl27H2O 0.1 g; and 100 ml of the stock solution (0.5 mg pyrene/1.0 ml acetone). For
preparation of agar plates, 15 g/l of agar was added. The final pH of the medium was adjusted to
7.0 with 1 N HCl and the medium was sterilized (121C for 20 min) prior to the addition of
pyrene. The prepared CFMM includes pyrene as only one carbon source. The fresh-broth culture
of P. vulgaris 4Bi in TSB (24 h culture at 30C) was spreaded aseptically on CFMM plates
supplemented with pyrene as the source of carbon and energy. The stock solution (0.5 mg/ml) of
pyrene was made in acetone and was sterilized by millipore micro syringe filter (0.45 m pore
size). Pyrene was purchased from Sigma (purity 95%). Acetone solution of pyrene was added to
CFMM agar and equal volume of acetone was used for control in dublicate. The inoculated
plates were sealed with parafilm to prevent evaporation. After 72 h of cultivation (at 30C),
colonies of P. vulgaris 4Bi were appeared on plate media. The pyrene utilization on CFMM
plates by colonies was observed as clear zones (haloes) around them (Figure 1). With
observation of growth and clear zones on the pyrene agar-plates, we suspected that P. vulgaris
4Bi was a bacterium degrading pyrene.
Marine Pollution Bulletin 57 (2008) 538543
Modified sublimation to isolate phenanthrene-degrading bacteria of the genera
Sphingomonas and Burkholderia from Xiamen oil port
X. Huang a,b,1, Y. Tian b,1, Y.R. Luo b, H.J. Liu b, Wei Zheng b, T.L. Zheng a,b,*
One milliliter of mixed culture was 10-fold serially diluted in sterile distilled water after
subculturing 10 times. Appropriate dilutions were plated onto nutritional agarnplates using the
spread plate method. The plates were incubated at 30 _C for 24 h before sublimation. Thirty
milliliters of phenanthrene stock solution was added to a 500 mL beaker. After the CH 2Cl2 had
evaporated completely, an inoculated agar plate was placed upside-down on the beaker, and the
joint between beaker and plate sealed with parafilm. The above-mentioned steps were carried out
under axenic conditions. Another disk filled with ice was then placed on the inverted plate. The
sublimation was performed at 95oC in a water bath for approximately 5 min to deposit a
phenanthrene layer on the agar. The plates were then incubated at 30 _C for 5 days. Different
bacteria were isolated from the agar plates and, particularly those with a clearing zone, were
recorded and preserved. Phenanthrene, purchased from SigmaAldrich, was
used as the sole carbon source for enrichment of degrading- bacteria. Phenanthrene stock
solution was freshly prepared in dichloromethane (CH2Cl2) at a concentration of 20 mg mL-1

Helix Vol. 2:105-111 (2012)

Identification and Isolation of Hydrocarbon Degrading Bacteria by
Molecular Characterization
1Jyothi K, 2K Surendra Babu*, 3Nancy Clara. K, 4Amita Kashyap
Determination of Bacterial Biodegradative
Activity by Turbidometry:
Turbidometry is to determine the bacterial growth by utilizing the hydrocarbons (1% petrol and
diesel given as carbon source in MSM broth. This shows whether the bacterium possess the
degrading activity of hydrocarbons like phenol, petrol and diesel. The degrading activities of
each isolates were obtained by using Mineral salt broth (MSB) in which 1% of each hydrocarbon
(petrol and diesel) was added and incubated at room temperature for 15 days. The growth of the
bacterium was measured by taking the O.D readings at 595nm from 0 hrs - 15 days at regular
intervals of 2 days against mineral salt medium as blank.