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Applied and Environmental Microbiology, November 2002, p. 5625-5633, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5625-5633.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Bacteria Belonging to the Genus Cycloclasticus Play a Primary Role in the Degradation of Aromatic Hydrocarbons Released in a Marine Environment

Yuki Kasai,^* Hideo Kishira, and Shigeaki Harayama

Marine Biotechnology Institute, 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan

Received 8 March 2002/ Accepted 16 August 2002

To identify the bacteria that play a major role in the aerobic degradation of petroleum polynuclear aromatic hydrocarbons (PAHs) in a marine environment, bacteria were enriched from seawater by using 2-methylnaphthalene, phenanthrene, or anthracene as a carbon and energy source. We found that members of the genus Cycloclasticus became predominant in the enrichment cultures. The Cycloclasticus strains isolated in this study could grow on crude oil and degraded PAH components of crude oil, including unsubstituted and substituted naphthalenes, dibenzothiophenes, phenanthrenes, and fluorenes. To deduce the role of Cycloclasticus strains in a coastal zone oil spill, propagation of this bacterial group on oil-coated grains of gravel immersed in seawater was investigated in beach-simulating tanks that were 1 m wide by 1.5 m long by 1 m high. The tanks were two-thirds filled with gravel, and seawater was continuously introduced into the tanks; the water level was varied between 30 cm above and 30 cm below the surface of the gravel layer to simulate a 12-h tidal cycle. The number of Cycloclasticus cells associated with the grains was on the order of 10³ cells/g of grains before crude oil was added to the tanks and increased to 3 x 10⁶ cells/g of grains after crude oil was added. The number increased further after 14 days to 10⁸ cells/g of grains when nitrogen and phosphorus fertilizers were added, while the number remained 3 x 10⁶ cells/g of grains when no fertilizers were added. PAH degradation proceeded parallel with the growth of Cycloclasticus cells on the surfaces of the oil-polluted grains of gravel. These observations suggest that bacteria belonging to the genus Cycloclasticus play an important role in the degradation of petroleum PAHs in a marine environment.

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* Corresponding author. Mailing address: Marine Biotechnology Institute, 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan. Phone: 81-193-26-6544. Fax: 81-193-26-6592. E-mail: yuki.kasai{at}mbio.jp.

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Applied and Environmental Microbiology, November 2002, p. 5625-5633, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5625-5633.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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